Anti-apoe antibodies

ABSTRACT

The present disclosure encompasses compositions and methods for effectively treating at least one symptom or sign of A plaque or cerebral amyloid angiopathy (CAA) associated symptoms, or for decreasing amyloid plaque load or CAA load. The method comprises administering an effective amount of an anti-ApoE antibody to a mammalian subject, such as to a human.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the priority of U.S. provisional application No.62/414,413, filed Oct. 28, 2016, and U.S. provisional application No.62/533,336 filed Jul. 17, 2017, each of which is hereby incorporated byreference in its entirety.

GOVERNMENTAL RIGHTS

This invention was made with government support under R01AG047644awarded by the National Institutes of Health. The government has certainrights in the invention.

FIELD OF THE INVENTION

The invention relates to anti-ApoE antibodies and compositions thereof.The invention also relates to compositions and methods for delayingand/or preventing Aβ amyloidosis. The invention also relates tocompositions and methods for delaying and/or preventing Aβ plaqueassociated symptoms and/or cerebral amyloid angiopathy (CAA) associatedsymptoms, such as those associated with Alzheimer's disease (AD) or CAAin a subject. In particular, the invention relates to modulating theconcentration of amyloid-β (Aβ) in the brain of a subject.

REFERENCE TO SEQUENCE LISTING

A paper copy of the sequence listing and a computer readable form of thesame sequence listing are appended below and herein incorporated byreference. The information recorded in computer readable form isidentical to the written sequence listing, according to 37 C.F.R.1.821(f).

BACKGROUND OF THE INVENTION

Alzheimer's disease (AD) is the most common cause of dementia and is anincreasing public health problem. It is currently estimated to afflictover 5 million people in the United States, with an expected increase to13 million by the year 2050. Alzheimer's disease leads to loss ofmemory, cognitive function, and ultimately loss of independence. Ittakes a heavy personal and financial toll on the subject and the family.Because of the severity and increasing prevalence of the disease in thepopulation, it is urgent that better treatments be developed.

Cerebral amyloid angiopathy (CAA) occurs in about 90% of individuals whodevelop AD, as well as in some individuals independently of AD. CAA canlead to ischemic and hemorrhagic strokes causing severe disability ordeath. There are no current treatments for CAA.

Biochemical, genetic, and animal model evidence implicates amyloid-β(Aβ) as a pathogenic peptide in AD and in most cases of CAA. Theneuropathologic and neurochemical hallmarks of AD include synaptic lossand selective neuronal death, a decrease in certain neurotransmitters,and the presence of abnormal proteinaceous deposits within neurons(neurofibrillary tangles) and in the extracellular space(cerebrovascular, diffuse, and neuritic plaques). The characteristicfeatures of CAA include the buildup of fibrillar forms of Aβ inpenetrating and leptomeningeal arterioles on the surface of the cerebralcortex. CAA can lead to ischemic or hemorrhagic stroke. The mainconstituent of the plaques seen in AD and CAA is Aβ, a 38-43 amino acidsequence peptide cleaved from the amyloid precursor protein (APP).

Throughout life, soluble Aβ is secreted primarily by neurons, but alsoother cell types. Excessive Aβ deposition may result from increased Aβsynthesis, e.g. as occurs in familial early-onset AD and in some casesof familial early onset CAA, decreased Aβ clearance in the brain, orincreased Aβ fibrillogenesis. The lack of compelling evidence that Aβover-production occurs in the more common late-onset forms of ADsuggests that insufficient Aβ clearance may drive Aβ deposition andamyloid plaque formation and CAA as well.

The apolipoprotein E gene (ApoE) remains the most widely replicatedgenetic risk factor for late-onset AD and CAA, with carriers of the ε4allele having a 3-15-fold greater risk as well as an earlier age ofdisease onset. ApoE4 carriers represent over 60% of the AD populationwhereas the ε2 allele is least represented in AD and may be protectivein some populations. The human ApoE isoforms differ at amino acidposition 112 or 158 (ε2 has Cys112, Cys158; ε3 has Cys112, Arg158; ε4has Arg112, Arg158). Mouse ApoE and human ApoE share ˜70% amino acidhomology. The amino acid differences within human ApoE isoforms haveeffects on the structure and stability of the protein. For example,ApoE4 has a greater propensity towards an unstructured, molten globulestate and is more likely to form aggregates, as compared to ApoE2 andApoE3.

In the brain, ApoE is mainly produced by glia and predominantlyfunctions to distribute cholesterol and lipids to neurons. The majorityof ApoE in the CNS is found on HDL-like lipoprotein particles, andlipidation of ApoE is regulated by the ABCA1 and ABCG1 transporters.ABCA1 knockout mice crossed to transgenic AD mice produce ApoE that ispoorly-lipidated and these mice have increased Aβ plaques. Conversely,the brains of AD transgenic mice overexpressing ABCA1 containwell-lipidated apoE lipoprotein particles and have reduced Asp plaquedeposition. Binding of ApoE to LDLR, LRP1, apoER2, and VLDLR, which areexpressed on various neural cell types, enables the uptake oflipoprotein particles through receptor-mediated endocytosis, processingof the lipid cargo, and recycling of the unbound receptors. The aminoacid difference at position 158 for ApoE2 confers decreased receptorbinding to the LDLR and humans with the ε2/ε2 genotype have increasedrisk for type III hyperlipoproteinemia.

The deposition of Aβ plaques in AD patients is ApoE isoform-dependent(E4>E3>E2), the extent of interaction between ApoE and Aβ and thefunction of their interaction remains controversial. Strong evidencesuggests the differential Aβ deposition is secondary to differences inapoE isoform-dependent Aβ clearance and aggregation. The clearance of Aβmonomers from the brains of AD transgenic mice is also ApoE isoformdependent (E2>E3>E4), and recent studies suggest ApoE4 may potentiallyhinder clearance by competing for binding to receptors that alsofunction to clear Aβ. The extent of ApoE lipidation may also affect theformation of Aβ oligomers with greater levels in ε4/ε4 AD patients thanε3/ε3. The present disclosure describes the discovery that anti-ApoEantibodies with certain characteristics provide a treatment for subjectswith Aβ amyloidosis.

SUMMARY OF THE INVENTION

One aspect of the invention encompasses an isolated anti-ApoE antibodycomprising (a) a light chain variable region comprising an L1 of SEQ IDNO: 86, an L2 of SEQ ID NO: 30, an L3 comprising SEQ ID NO: 88, or anycombination thereof; and/or (b) a heavy chain variable region comprisingan H1 comprising SEQ ID NO: 90, an H2 comprising SEQ ID NO: 92, an H3comprising SEQ ID NO: 94, or any combination thereof. Compositionscomprising the antibody, including but not limited to pharmaceuticalcompositions, are contemplated herein. In certain embodiments theantibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 87, an L2 of SEQ ID NO: 30, an L3 comprising SEQ ID NO:89, or any combination thereof; and/or (b) a heavy chain variable regioncomprising an H1 comprising SEQ ID NO: 91, an H2 comprising SEQ ID NO:93, an H3 comprising SEQ ID NO: 95, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 78, an L2 of SEQ ID NO: 24, an L3 comprising SEQ ID NO:25, or any combination thereof; and/or (b) a heavy chain variable regioncomprising an H1 comprising SEQ ID NO: 80, an H2 comprising SEQ ID NO:82, an H3 comprising SEQ ID NO: 84, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 79, an L2 of SEQ ID NO: 24, an L3 comprising SEQ ID NO:25, or any combination thereof; and/or (b) a heavy chain variable regioncomprising an H1 comprising SEQ ID NO: 81, an H2 comprising SEQ ID NO:83, an H3 comprising SEQ ID NO: 86, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 105, an L2 of SEQ ID NO: 106, an L3 comprising SEQ ID NO:107, or any combination thereof; and/or (b) a heavy chain variableregion comprising an H1 comprising SEQ ID NO: 109, an H2 comprising SEQID NO: 111, an H3 comprising SEQ ID NO: 113, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 105, an L2 of SEQ ID NO: 106, an L3 comprising SEQ ID NO:108, or any combination thereof; and/or (b) a heavy chain variableregion comprising an H1 comprising SEQ ID NO: 110, an H2 comprising SEQID NO: 112, an H3 comprising SEQ ID NO: 114, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 23, an L2 of SEQ ID NO: 24, an L3 comprising SEQ ID NO:25, or any combination thereof; and/or (b) a heavy chain variable regioncomprising an H1 comprising SEQ ID NO: 26, an H2 comprising SEQ ID NO:27, an H3 comprising SEQ ID NO: 28, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 29, an L2 of SEQ ID NO: 30, an L3 comprising SEQ ID NO:31, or any combination thereof; and/or (b) a heavy chain variable regioncomprising an H1 comprising SEQ ID NO: 32, an H2 comprising SEQ ID NO:33, an H3 comprising SEQ ID NO: 34, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 47, an L2 of SEQ ID NO: 24, an L3 comprising SEQ ID NO:25, or any combination thereof; and/or (b) a heavy chain variable regioncomprising an H1 comprising SEQ ID NO: 48, an H2 comprising SEQ ID NO:49, an H3 comprising SEQ ID NO: 50, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 51, an L2 of SEQ ID NO: 24, an L3 comprising SEQ ID NO:25, or any combination thereof; and/or (b) a heavy chain variable regioncomprising an H1 comprising SEQ ID NO: 52, an H2 comprising SEQ ID NO:53, an H3 comprising SEQ ID NO: 54, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 55, an L2 of SEQ ID NO: 24, an L3 comprising SEQ ID NO:25, or any combination thereof; and/or (b) a heavy chain variable regioncomprising an H1 comprising SEQ ID NO: 56, an H2 comprising SEQ ID NO:57, an H3 comprising SEQ ID NO: 58, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 59, an L2 of SEQ ID NO: 24, an L3 comprising SEQ ID NO:25, or any combination thereof; and/or (b) a heavy chain variable regioncomprising an H1 comprising SEQ ID NO: 60, an H2 comprising SEQ ID NO:61, an H3 comprising SEQ ID NO: 62, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 63, an L2 of SEQ ID NO: 30, an L3 comprising SEQ ID NO:64, or any combination thereof; and/or (b) a heavy chain variable regioncomprising an H1 comprising SEQ ID NO: 65, an H2 comprising SEQ ID NO:66, an H3 comprising SEQ ID NO: 67, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 68, an L2 of SEQ ID NO: 24, an L3 comprising SEQ ID NO:25, or any combination thereof; and/or (b) a heavy chain variable regioncomprising an H1 comprising SEQ ID NO: 69, an H2 comprising SEQ ID NO:70, an H3 comprising SEQ ID NO: 71, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 72, an L2 of SEQ ID NO: 73, an L3 comprising SEQ ID NO:74, or any combination thereof; and/or (b) a heavy chain variable regioncomprising an H1 comprising SEQ ID NO: 75, an H2 comprising SEQ ID NO:76, an H3 comprising SEQ ID NO: 77, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 105, an L2 of SEQ ID NO: 106, an L3 comprising SEQ ID NO:123, or any combination thereof; and/or (b) a heavy chain variableregion comprising an H1 comprising SEQ ID NO: 124, an H2 comprising SEQID NO: 125, an H3 comprising SEQ ID NO: 126, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 99, an L2 of SEQ ID NO: 100, an L3 comprising SEQ ID NO:101, or any combination thereof; and/or (b) a heavy chain variableregion comprising an H1 comprising SEQ ID NO: 102, an H2 comprising SEQID NO: 103, an H3 comprising SEQ ID NO: 104, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses an isolated anti-ApoEantibody comprising (a) a light chain variable region comprising an L1of SEQ ID NO: 105, an L2 of SEQ ID NO: 106, an L3 comprising SEQ ID NO:117, or any combination thereof; and/or (b) a heavy chain variableregion comprising an H1 comprising SEQ ID NO: 118, an H2 comprising SEQID NO: 119, an H3 comprising SEQ ID NO: 120, or any combination thereof.Compositions comprising the antibody, including but not limited topharmaceutical compositions, are contemplated herein. In certainembodiments the antibody is a humanized antibody.

Another aspect of the invention encompasses a method of improving aclinical sign of Aβ amyloidosis which comprises administering aneffective amount of an anti-ApoE antibody to a living human subject. Inanother aspect, the invention encompasses a method of effectivelytreating at least one clinically detectable Aβ plaque associated symptomand/or CAA associated symptom which comprises administering an effectiveamount of an anti-ApoE antibody to a living human subject. In anotheraspect, the invention encompasses a method of treating Alzheimer'sdisease which comprises administering an effective amount of ananti-ApoE antibody to a living human subject. In another aspect, theinvention encompasses a method of treating CAA which comprisesadministering an effective amount of an anti-ApoE antibody to a livinghuman subject. In each of the above aspects, the anti-ApoE antibody maybe an antibody described above.

Yet another aspect of the invention encompasses a pharmaceuticalcomposition useful to treat at least one clinically detectable Aβ plaqueassociated symptom. The composition comprises a pharmaceuticallyeffective amount of an anti-ApoE antibody adapted for administration toa living human subject. The anti-ApoE antibody may be an antibodydescribed above. In an aspect, the medicinal composition is effectivelyadministered to a living subject systemically.

Yet another aspect of the invention encompasses a pharmaceuticalcomposition useful to treat at least one clinically detectable CAAassociated symptom. The composition comprises a pharmaceuticallyeffective amount of an anti-ApoE antibody adapted for administration toa living human subject. The anti-ApoE antibody may be an antibodydescribed above. In an aspect, the pharmaceutical composition iseffectively administered to a living subject systemically.

Yet another aspect of the invention encompasses a pharmaceuticalcomposition useful to treat Alzheimer's disease. The compositioncomprises a pharmaceutically effective amount of an anti-ApoE antibodyadapted for administration to a living human subject. The anti-ApoEantibody may be an antibody described above. In an aspect, thepharmaceutical composition is effectively administered to a livingsubject systemically.

Yet another aspect of the invention encompasses a pharmaceuticalcomposition useful to treat CAA. The composition comprises apharmaceutically effective amount of an anti-ApoE antibody adapted foradministration to a living human subject. The anti-ApoE antibody may bean antibody described above. In an aspect, the pharmaceuticalcomposition is effectively administered to a living subjectsystemically.

Still another aspect the invention encompasses a kit comprising acontainer containing a pharmaceutical composition comprising apharmaceutically effective amount of an anti-ApoE antibody adapted foradministration to a living human subject and any medical devices to beused for said administration. The anti-ApoE antibody may be an antibodydescribed above.

Other aspects and iterations of the invention are detailed below.

BRIEF DESCRIPTION OF THE FIGURES

The application file contains at least one photograph executed in color.Copies of this patent application publication with color photographswill be provided by the Office upon request and payment of the necessaryfee.

FIG. 1 depicts an image of a Western blot. The brain lysates from ApoEKO mice or mice expressing ApoE3, ApoE4 or murine ApoE wereimmunoblotted with GA-50, HJ151, or HJ152. The Western blot shows thatHJ151 is ApoE4 specific and HJ152 recognizes both ApoE3 and ApoE4. GA-50is a positive control.

FIG. 2 depicts an image of a Western blot. The brain lysates from ApoEKO mice or mice expressing ApoE3, ApoE4 or murine ApoE wereimmunoblotted with HJ153 or HJ154. The Western blot shows that HJ153 andHJ154 recognize both ApoE3 and ApoE4.

FIG. 3 depicts images of brain tissue from 5XFAD APP transgenic miceexpressing different human ApoE isoforms and stained using biotinylatedHJ153 or HJ154 antibodies. The figure shows these two antibodies stainApoE in the neuropil, in astrocytes, and in amyloid plaques if plaquesare present. Brain tissue from ApoE KO mouse was used as negativecontrol.

FIG. 4 depicts images of several immunoblots. Samples containing ApoE2,ApoE3 or ApoE4 were immunoprecipitated using HJ152, HJ153 or HJ154.Materials immunoprecipitated with anti-ApoE antibodies (labeled as IP)and the solution after immunoprecipitation (labeled as post IP) wereimmunoblotted after running an SDS-PAGE gel and transfer tonitrocellulose membrane using GA-50, a commercial anti-ApoE antibody.WUE4, a monoclonal antibody against human ApoE, was used as a positivecontrol. The results showed that HJ152 was able to immunoprecipitatesome of the ApoE from the samples while there was still some ApoEremaining in the post IP product. HJ153 and HJ154 were able toimmunoprecipitate all ApoE from the samples.

FIG. 5A-H depicts graphs showing aspects of various antibodies.Specifically, FIG. 5A-D graphically shows the results of an ELISA usingvarious antibodies. ELISA plates were coated with 0.5 μg/ml ofrecombinant apoE2 (square), apoE3 (open circle), or apoE4 (closedcircle). Then different concentrations of (A) HJ151, (B) HJ153, (C)HJ154, or (D) HJ156 were loaded on the plates. Horse-radish peroxidaselabeled goat anti-mouse secondary antibodies were used to detectbinding. The results show that HJ153 and HJ154 bind ApoE2, ApoE3 andApoE4 as detected in the ELISA. HJ151 is specific for ApoE4 and HJ156binds only ApoE3 and ApoE4. FIG. 5E-H are graphs depicting surfaceplasmon resonance profiles for various antibodies. Anti-ApoE antibodieswere serially diluted 3-fold (starting at 100 nM for HJ153 (E), and 1000nM for HJ151 (F), and HJ1156 (G)) for detection of binding tobiotinylated-recombinant apoE4 captured on a streptavidin chip. Sampleswere injected at a flow rate of 30 μl/minute. (H) Apparent KD values ofHJ151, HJ153 and HJ156 were calculated based on the SPR experiment.

FIG. 6 depicts a graph showing some anti-ApoE antibodies decrease Aβplaques in APP/PS1-21 E4/E4 mice after ICV infusion. APP/PS1-21 E4/E4mice received continuous intracerebroventricular (ICV) infusion of PBS(negative control), mouse IgG2ab (negative control), HJ5.1 (anti-Aβantibody, positive control), or anti-apoE antibody (HJ151, HJ154, andHJ156) beginning at 2 months of age. Anti-ApoE antibody or controlantibody (2 mg/ml) was filled into a subcutaneous osmotic minipump(Alzet, model 2006) and infused through a surgically implanted catheterinto the left lateral cerebral ventricle (bregma −0.4 mm, 1.0 mm lateralto midline, 2.5 mm below the skull), infusing fluid at the speed of 1.2μl/min for 6 weeks. At the age of 3.5-months, the mice were perfused andthe sections were stained for Aβ plaques using anti-Aβ antibody HJ3.4B.The percent of area covered by plaques in the cerebral cortex dorsal tohippocampus was quantified. One-way ANOVA followed by Tukey post-testwas used to analyze the data (*, p<0.05; **, p<0.01; ***, p<0.001;n=10-12/group).

FIG. 7 depicts a graph showing some anti-ApoE antibodies decreasefibrillar plaque load after ICV infusion in APP/PS1-21 E4/E4 mice.APP/PS1-21 E4/E4 mice received continuous intracerebroventricular (ICV)infusion of PBS (negative control), mouse IgG2ab (negative control),HJ5.1 (anti-Aβ antibody, positive control), or anti-apoE antibody(HJ151, HJ154, and HJ156) from 2 months of age. Anti-apoE antibody orcontrol antibody (2 mg/ml) was filled into a subcutaneous osmoticminipump (Alzet, model 2006) and infused through a surgically implantedcatheter into the left lateral cerebral ventricle (bregma −0.4 mm, 1.0mm lateral to midline, 2.5 mm below the skull) at the speed of 1.2μl/min for 6 weeks. At the age of 3.5-months, the mice were perfused andthe brain sections were stained for fibrillar plaques using ThioflavineS. The area covered by plaques in the cerebral cortex dorsal tohippocampus was quantified. One-way ANOVA followed by Tukey post-testwas used to analyze the data (*, p<0.05; n=10-12/group).

FIG. 8A-F depicts graphs showing that anti-ApoE antibodies decreaseinsoluble Aβ42 in APP/PS1-21 E4/E4 mice after ICV infusion. APP/PS1-21E4/E4 mice received continuous intracerebroventricular (ICV) infusion ofPBS (negative control), mouse IgG2ab (negative control), HJ5.1 (anti-Aβantibody, positive control), or anti-apoE antibody (HJ151, HJ154, andHJ156) from 2 months of age. Anti-apoE antibody or control antibody (2mg/ml) was filled into a subcutaneous osmotic minipump (Alzet, model2006) and infused through a surgically implanted catheter into the leftlateral cerebral ventricle (bregma −0.4 mm, 1.0 mm lateral to midline,2.5 mm below the skull) at the speed of 1.2 μl/min for 6 weeks(n=10-12/group). At the age of 3.5-months, the mice were perfused andthe cortical tissue was homogenized in PBS, Triton and 5M Guanidinesequentially. The Aβ levels in soluble (A, D) and insoluble (B, C, E, F)fractions were measured (Aβ40 data shown in A-C, Aβ42 data shown inD-F). Student's t-test was used for data analysis. Anti-apoE antibodiesreduced Aβ₄₂ levels in different fractions (*, p<0.05; **, p<0.01).

FIG. 9 depicts a graph showing the body weight of APP/PS1-21 E4/E4 miceintraperitoneally injected with anti-ApoE antibodies (1 injection/weekfor 7 weeks).

FIG. 10 depicts a graph showing that HJ151, HJ155, and HJ156 do notsignificantly decrease soluble Aβ40 in the PBS fraction of brain tissuehomogenate in APP/PS1-21 E4/E4 mice intraperitoneally injected withanti-ApoE antibodies (1 injection/week for 7 weeks).

FIG. 11 depicts a graph showing that HJ151, HJ155, and HJ156 do notsignificantly decrease soluble Aβ42 in the PBS fraction of brain tissuehomogenate of APP/PS1-21 E4/E4 mice intraperitoneally injected withanti-ApoE antibodies (1 injection/week for 7 weeks).

FIG. 12 depicts a graph showing that HJ156 significantly decreasesinsoluble Aβ₄₀ in the 5M guanidine fraction of brain tissue homogenateof APP/PS1-21 E4/E4 mice intraperitoneally injected with anti-ApoEantibodies (1 injection/week for 7 weeks). (*p<0.05)

FIG. 13 depicts a graph showing that HJ156 significantly decreasesinsoluble Aβ₄₂ in the 5M guanidine fraction of brain tissue homogenateof APP/PS1-21 E4/E4 mice intraperitoneally injected with anti-ApoEantibodies (1 injection/week for 7 weeks). (*p<0.05, (**p<0.01).

FIG. 14 is a graph depicting relative antibody concentration in thecortex of APPPS1-21/APOE4 mice expressing recombinant (r) HJ151, r HJ151with D265A mutation (Δ), rHJ156, and rHJ156Δ. APPPS1-21/APOE4 mice wereinjected at PO with AAV2/8 that express full length rHJ151 and rHJ156with or without the D265A mutation. At the age of 3.5 months, antibodyconcentration in the PBS soluble fraction of cortex was measured byELISA. The relative level of each antibody was calculated by using itshybridoma-derived, purified antibody as a standard.

FIG. 15 depicts a graph showing that HJ151 and HJ156 significantlyreduce insoluble Aβ₄₂ in the guanidine fraction of brain tissuehomogenate of APP/PS1-21 E4/E4 mice injected with anti-ApoE antibodiesexpressed in the brain with the use of an adenoassociated virus (AAV)2/8 vector. HJ151 and HJ156 with a D265A mutation (HJ151Δ and HJ156Δ)have no effect.

FIG. 16 depicts a graph showing that HJ151 and HJ156 reduce insolubleAβ₄₀ in the guanidine fraction of brain tissue homogenate of APP/PS1-21E4/E4 mice injected with anti-ApoE antibodies expressed in the brainwith the use of an adenoassociated virus (AAV) 2/8 vector. HJ151 andHJ156 with a D265A mutation (HJ151Δ and HJ156Δ) have no effect.

FIG. 17A-B depict graphs showing that HJ151 and HJ156 significantlyreduce amyloid β (A) and fibrillar plaque area (B) in brain sectionsfrom APP/PS1-21 E4/E4 mice injected with anti-ApoE antibodies expressedin the brain with the use of an adenoassociated virus (AAV) 2/8 vector.HJ151 and HJ156 with a D265A mutation (HJ151Δ and HJ156Δ) have noeffect.

FIG. 18A-G depicts unfixed tissue sections (20 μm thickness) fromAPPPS1-21/apoE^(4/4) (APPPS1-21/E4), 5XFAD/apoE knockout (5XFAD/EKO)mice and human apoE^(4/4) (human E4) brains that were stained usingbiotinylated (“B”) antibodies HJ3.4B (A), HJ151B (B), HJ152B (C), HJ153B(D), HJ154B (E), HJ155B (F), and HJ156B (G).

FIG. 19A-D shows antibody detection of ApoE in human plasma lipoproteinparticles. (A) shows a schematic of a plasma binding assay used todemonstrate the detection of ApoE in lipoprotein particles from humanplasma coated onto an ELISA plate, while (B-D) each show a graph of theresults from the binding assay for various antibodies. Detection of ApoEis observed with HJ153, HJ154, HJ1510, HJ1511, HJ1517, HJ1530, andHJ1534.

FIG. 20A-C shows antibody detection of ApoE in a competition ELISA. (A)shows a schematic of a plasma competition assay used in assessingpreference for antibody binding to coated recombinant alipidated ApoE4(recApoE; 0.5 ug/mL) after pre-incubation in various dilutions of humanplasma, while (B-C) each show a graph of the results from competitionELISA for various antibodies. “x ApoE” is an abbreviation for “anti-ApoEantibody.”

FIG. 21A-D shows images of unfixed frozen brain sections from APP/PS1-21E2/E2 mice that were immunostained for ApoE with anti-ApoE antibodyHJ156B at 20 ug/ml (A,C) or 50 ug/ml (B,D) at two differentmagnifications. Scale bars, 400 μm.

FIG. 22A-D shows images of unfixed frozen brain sections from APP/PS1-21E3/E3 mice that were immunostained for ApoE with anti-ApoE antibodyHJ156B at 20 ug/ml (A,C) or 50 ug/ml (B,D) at two differentmagnifications. Scale bars, 400 μm.

FIG. 23A-D shows images of unfixed frozen brain sections from APP/PS1-21E4/E4 mice that were immunostained for ApoE with anti-ApoE antibodyHJ156B at 20 ug/ml (A,C) or 50 ug/ml (B,D) at two differentmagnifications. Scale bars, 400 μm.

FIG. 24A-D shows images of unfixed frozen brain sections from APP/PS1-21E4/E4 mice that were immunostained for ApoE with anti-ApoE msIgG/huFcchimeric antibody HJ153 and a biotinylated rabbit anti-human IgGsecondary antibody. Primary antibody dilutions of (A) 0.4 μg/mL, (B) 2μg/mL, (C) 20 μg/mL, and (D) 50 μg/mL were used. Scale bars, 400 μm.

FIG. 25A-D shows images of unfixed frozen brain sections from APP/PS1-21E4/E4 mice that were immunostained for ApoE with anti-ApoE msIgG/huFcchimeric antibody HJ156 and a biotinylated rabbit anti-human IgGsecondary antibody. Primary antibody dilutions of (A) 0.4 μg/mL, (B) 2μg/mL, (C) 20 μg/mL, and (D) 50 μg/mL were used. Scale bars, 400 μm.

FIG. 26A-D shows images of unfixed frozen brain sections from APP/PS1-21E4/E4 mice that were immunostained for ApoE with anti-ApoE msIgG/huFcchimeric antibody HJ1514 and a biotinylated rabbit anti-human IgGsecondary antibody. Primary antibody dilutions of (A) 0.4 μg/mL, (B) 2μg/mL, (C) 20 μg/mL, and (D) 50 μg/mL were used. Scale bars, 400 μm.

FIG. 27A-D shows images of unfixed frozen brain sections from APP/PS1-21E4/E4 mice that were immunostained for ApoE with anti-ApoE msIgG/huFcchimeric antibody HJ1518 and a biotinylated rabbit anti-human IgGsecondary antibody. Primary antibody dilutions of (A) 0.4 μg/mL, (B) 2μg/mL, (C) 20 μg/mL, and (D) 50 μg/mL were used. Scale bars, 400 μm.

FIG. 28A-D shows images of unfixed frozen brain sections from APP/PS1-21E4/E4 mice that were immunostained for ApoE with anti-ApoE msIgG/huFcchimeric antibodies at 2 μg/mL—(A) HJ153 chimeric, (B) HJ156 chimeric,(C) HJ1514 chimeric, (D) HJ1518 chimeric. Scale bars, 400 μm.

FIG. 29 shows a graph illustrating Aβ deposition, measured as % area Aβstaining, in APP/PS1-21 E4/E4 mice treated with weekly intra-peritoneal(IP) injections of PBS, HJ16.6 (IgG negative control), and the anti-ApoEantibodies HJ151, HJ155, and HJ156 from 2 months of age to 3.5 months ofage. All antibodies were administered at 50 mg/kg. Brain sections from 2month old untreated, and 3.5 month old treated mice were stained withanti-Aβ antibody HJ3.4-biotin. Aβ staining was quantified from cortex,n=10 per group. *p<0.05 compared to PBS and HJ16.6 groups.

FIG. 30 shows a graph illustrating Aβ plaque number in APP/PS1-21 E4/E4mice treated with weekly intra-peritoneal (IP) injections of PBS, HJ16.6(IgG negative control), and the anti-ApoE antibodies HJ151, HJ155, andHJ156 from 2 months of age to 3.5 months of age. All antibodies wereadministered at 50 mg/kg. Brain sections from 2 month old untreated, and3.5 month old treated mice were stained with anti-Aβ antibodyHJ3.4-biotin. Aβ plaque number, detected by Aβ plaque staining, wasquantified from cortex, n=10 per group. *p<0.05 compared to PBS andHJ16.6 groups.

FIG. 31 shows a graph illustrating fibrillar Aβ plaque deposition,measured as % area X-34 staining, in APP/PS1-21 E4/E4 mice treated withweekly intra-peritoneal (IP) injections of PBS, HJ16.6 (IgG negativecontrol), and the anti-ApoE antibodies HJ151, HJ155, and HJ156 from 2months of age to 3.5 months of age. All antibodies were administered at50 mg/kg. Brain sections from 2 month old untreated, and 3.5 month oldtreated mice were stained with X-34, a dye that stains fibrillarplaques. X-34 was quantified from cortex, n=10 per group. *p<0.05compared to PBS and HJ16.6 groups.

FIG. 32 shows a graph illustrating fibrillar Aβ plaque number inAPP/PS1-21 E4/E4 mice treated with weekly intra-peritoneal (IP)injections of PBS, HJ16.6 (IgG negative control), and the anti-ApoEantibodies HJ151, HJ155, and HJ156 from 2 months of age to 3.5 months ofage. All antibodies were administered at 50 mg/kg. Brain sections from 2month old untreated, and 3.5 month old treated mice were stained withX-34, a dye that stains fibrillar plaques. Fibrillar Aβ plaque number,detected by X-34 staining, was quantified from cortex, n=10 per group.*p<0.05 compared to PBS and HJ16.6 groups.

FIG. 33A-F shows images of brain sections immunostained for Aβ withanti-Aβ antibody (HJ3.4-biotin). (A,D) Brain sections from non-treated,2 month old mice before the development of Aβ plaques. (B,E) Brainsections from 3.5 month old mouse treated with PBS. (C,F) Brain sectionsfrom 3.5 month old mouse treated with negative control IgG (HJ16.6) at50 mg/kg. Scale bar 300 μm, n=10 per group.

FIG. 34A-F shows images of brain sections immunostained for Aβ withanti-Aβ antibody (HJ3.4-biotin). (A,D) Brain sections from 3.5 month oldmouse treated with HJ151 at 50 mg/kg. (B,E) Brain sections from 3.5month old mouse treated with HJ155 at 50 mg/kg. (C,F) Brain sectionsfrom 3.5 month old mouse treated with HJ156 at 50 mg/kg. Scale bar 300μm, n=10 per group.

FIG. 35A-F shows images of brain sections stained with X-34 dye thatrecognizes fibrillar plaques. (A,D) Brain sections from non-treated, 2month old mice before the development of Aβ plaques. (B,E) Brainsections from 3.5 month old mouse treated with PBS. (C,F) Brain sectionsfrom 3.5 month old mouse treated with negative control IgG (HJ16.6) at50 mg/kg. Scale bar 300 μm, n=10 per group.

FIG. 36A-F shows images of brain sections stained with X-34 dye thatrecognizes fibrillar plaques. (A,D) Brain sections from 3.5 month oldmouse treated with HJ151 at 50 mg/kg. (B,E) Brain sections from 3.5month old mouse treated with HJ155 at 50 mg/kg. (C,F) Brain sectionsfrom 3.5 month old mouse treated with HJ156 at 50 mg/kg. Scale bar 300μm, n=10 per group.

FIG. 37A-F are graphs depicting the binding profile of HJ151, HJ153 andHJ156 with lipidated apoE. (A,B) ELISA following coating of plasma onplates. Plasma from apoE KO, APOE2, APOE3 and APOE4 mice was coated ontothe plates. Chi153 and chi156 of different concentrations were loaded.The captured antibodies were detected with HRP-goat anti-human IgGantibody. (C,D) ELISA following coating of antibodies on plates. Plateswere coated with HJ153 and HJ156. Plasma from mice with differentgenotypes was loaded. The captured ApoE was detected withHRP-goat-polyclonal anti-ApoE. (E) Plasma competition experiment wasperformed via ELISA format. Recombinant ApoE4 was coated onto theplates. HJ151 (50 nM), HJ153 (4 nM), and HJ156 (50 nM) pre-incubatedwith serially diluted plasma from APOE4 KI mice was loaded to theplates. The HJ15 antibodies bound to the plates were detected withHRP-Goat anti-mouse IgG antibodies. (F) Plasma antibody concentrationsof HJ153, HJ156, or control IgG following IP injection into APOE4 KI orEKO mice. HJ156 was dosed at 2, 10 and 50 mg/kg and plasma samplescollected by submandibular puncture. HJ153 was dosed at 10 mg/kg.Control murine IgG2a (msIgG2a) was anti-Her2 and dosed at 10 mg/kg.Quantification of dosed antibodies in plasma was by antigen captureELISA using coated recombinant apoE4 to detect HJ153 or HJ156 withrecombinant Her2 used to detect the control IgG. Peripheral clearance ofHJ156 is similar to Control IgG with target mediated clearance observedby day 14. HJ153 exhibited high clearance and reached lower limits by 48hours.

FIG. 38A-E are graphs depicting the binding of HJ151 and HJ156 withApoE4 in the amyloid plaques in unfixed mouse brain sections andspecificity for heat-induced aggregates of ApoE4. (A) Unfixed frozenbrain sections from APPPS1/APOE4 or APPPS1/APOE KO mice were stainedwith anti-Aβ antibody HJ3.4, anti-ApoE antibodies HJ151 and HJ156.(Scale bar=400 μm). (B) Binding of HJ151, HJ153 and HJ156 to untreatedrecombinant ApoE4 (untreated) and ApoE4 that has been incubated at 40°C. for 24 hr (40 C). (C) Incubation of ApoE4 at 40° C. for 24 hourresults in the formation of aggregates recovered in the pellet fractionafter ultracentrifugation at 186,000 g for 1 hour. 1: untreated ApoE4.2: ApoE4 that has been incubated at 40° C. for 24 hr. Supernatant (S)and pellet (P) from ultracentrifugation were resolved on SDS-PAGE andstained by Coomassie blue. (D) Binding of HJ156 to differentpreparations of ApoE4 loaded at the same concentration on the ELISAplate. Sup of untreated: Supernatant fraction of untreated ApoE4 fromultracentrifugation. Sup/Pel of 24 h 40 C: Supernatant/pellet fractionof ApoE4 incubated at 40° C. for 24 hour. (E) Binding of HJ156 tountreated and 40° C. incubated ApoE4 with and without denaturation by 1%SDS or 4M guanidine HCl.

FIG. 39A-D are images of HJ151, HJ156 and control antibody binding tohuman ApoE4 in living mouse brain. (A) Negative control IgG2ab (B) HJ151and (C) HJ156 conjugated with Alexa 594 were applied directly onto thesurface of the brain in living APPPS1-21/APOE4 mice that were 6 monthsof age and the binding of antibodies was observed under 2-photonmicroscopy. The amyloid was labeled using methoxy-X04. The middle panelis the higher power images of the area in the white frame of the toppanel. Arrows indicate plaques. The bottom panel is the higher powerimages of the area in the yellow frame of the top panel. Arrows indicateCAA. (D) Control huIgG or chi156 at 50 mg/kg body weight (i.p.) wereinjected for one (0 hour) or two doses (0 and 48 hour) andAPPPS1-21/APOE4 mice sacrificed at 48 hours after final injection. Theantibodies in the brain were detected by biotinylated rabbit anti-humanIgG followed by DAB. (Left panel: Bar=1 mm. right panel: high powerimage of the indicated areas in the left panel. Bar=300 μm)

FIG. 40 depicts a graph showing that HJ156 activates microglia tofacilitate plaque clearance. 4-month old APPPS1-21/APOE4 mice thatalready had existing plaques were administered a short-term treatment ofHJ151 and HJ156 antibodies (4 doses by IP injection every 3 days).Stained sections were analyzed for CD45+ microglia area relative to theamount of fibrillar plaques. HJ151 had no effect on the amount ofactivated microglia. *p<0.05.

FIG. 41 depicts staining of microglia and plaques after acuteimmunization of HJ156. At the age of 4 months, the APPPS1/APOE4 micereceived 4 IP injections of 50 mg/kg weight of antibodies every threedays. The mice were sacrificed 24 hours after the final injection andthe plaques were stained with X34 and the activated microglia wasstained with CD45 (Bar=1 mm).

FIG. 42A-B graphically shows the results of a chronic dose-rangeefficacy study. Two-month-old APPPS1-21/APOE4 mice were IP injected witha weekly dose of IgG2ab (50 mg/kg), anti-Aβ HJ3.4 (50 mg/kg) or HJ156(2, 10 and 50 mg/kg) for 12 doses (n=17-18/group, mixed gender). A groupof 2 months old animals (n=12) were harvested to determine the baselinelevel of Aβ. At the age of 4.5 months (3 days after the last dose), themice were perfused with ice-cold PBS containing 0.3% heparin. Thecerebral cortices were sequentially homogenized with cold PBS and 5 Mguanidine buffer in the presence of 1× protease inhibitor mixture. (A)Aβ40 and (B) Aβ42 in the guanidine fraction were determined by ELISA.One-way ANOVA followed by Tukey post-test was performed to comparedifferent groups. Data were expressed as mean±SEM. *p<0.05, **p<0.01,***p<0.001.

FIG. 43A-C graphically shows the binding of (A) HJ151, (B) HJ154, and(C) HJ156 antibodies to untreated recombinant and heat-inducedaggregates of ApoE2, ApoE3, and ApoE4. The aggregated ApoE was inducedby incubating ApoE at 1 mg/ml concentration at 40° C. for 24 hours. Theaggregates then were recovered in the pellet fraction followingultracentrifugation at 186,000 g for 1 hour. For ELISA, the untreatedApoE or heat-induced aggregates of ApoE were coated directly to ELISAplates at 0.5 μg/ml overnight at 4° C. After three washes with PBS, thewells were blocked with 1% BSA-PBS for 1 hour at room temperature withshaking at 500 rpm. The blocked wells were washed once with PBS andsubsequently loaded with antibodies at serial concentrations (startingat 100 nM with 5-fold dilutions thereafter). Bound antibodies weredetected with HRP-labeled goat anti-mouse IgG (Jackson Immunoresearch)and visualized with TMB substrate at OD₆₅₀ (reaction stopped with BioFXstop solution). The results show that HJ151 only detected ApoE4 and hadsome preference for heat-treated ApoE4, HJ154 detected all threeisoforms of ApoE and had no preference for conformation, and HJ156detected both ApoE3 and ApoE4 and preferred heat treated ApoE.

DETAILED DESCRIPTION

Applicants have discovered anti-ApoE antibodies and methods of using theanti-ApoE antibodies to treat Aβ amyloidosis. The method compriseseffectively administering to a living subject a therapeuticallyeffective amount of an anti-ApoE antibody that (a) binds to human ApoE4with a KD between about 0.1 pM to about 10 μM, or between about 0.1 pMto about 1 μM, (b) preferentially binds recombinant alipidated humanApoE4 as compared to ApoE4 derived from human plasma or human ApoE4derived from plasma of a transgenic mouse expressing human ApoE4, and(c) binds to human ApoE in amyloid plaque in unfixed brain tissue. Thepresent invention encompasses the discovery that anti-ApoE antibodieswith these characteristics provide a treatment for subjects with Aβamyloidosis including, but not limited to, subjects diagnosed with adisease characterized by brain Aβ plaques, subjects diagnosed with adisease characterized by vascular Aβ plaques in the brain, subjectsdiagnosed with Aβ plaque-associated symptoms, subjects diagnosed withCAA-associated symptoms, subjects with clinical signs of Aβ amyloidosisthat may or may not have Aβ plaque associated symptoms and/or CAAassociated symptoms, subjects diagnosed with Alzheimer's disease, andsubjects diagnosed with CAA (collectively referred to, herein, as“subjects in need of treatment”). Methods for identifying clinical signsof Aβ amyloidosis in asymptomatic patients are known in the art anddiscussed below.

I. Definitions

The term “subject” refers to a human, or to a non-human animalexpressing human ApoE.

The terms “treat,” “treating,” or “treatment” as used herein, refer toboth therapeutic treatment and prophylactic or preventative measures,wherein the object is to prevent or slow down (lessen) an undesiredphysiological change or disease/disorder. Beneficial or desired clinicalresults include, but are not limited to, alleviation of symptoms,diminishment of extent of disease, stabilized (i.e., not worsening)state of disease, a delay or slowing of disease progression,amelioration or palliation of the disease state, and remission (whetherpartial or total), whether detectable or undetectable. “Treatment” canalso mean prolonging survival as compared to expected survival if notreceiving treatment. Those in need of treatment include those alreadywith the disease, condition, or disorder as well as those prone to havethe disease, condition or disorder or those in which the disease,condition or disorder is to be prevented.

The term “Aβ” refers to peptides derived from a region in the carboxyterminus of a larger protein called amyloid precursor protein (APP). Thegene encoding APP is located on chromosome 21. There are many forms ofAβ that may have toxic effects: Aβ peptides are typically 37-43 aminoacid sequences long, though they can have truncations and modificationschanging their overall size. They can be found in soluble and insolublecompartments, in monomeric, oligomeric and aggregated forms,intracellularly or extracellularly, and may be complexed with otherproteins or molecules. The adverse or toxic effects of Aβ may beattributable to any or all of the above noted forms, as well as toothers not described specifically. For example, two such Aβ isoformsinclude Aβ40 and Aβ42; with the Aβ42 isoform being particularlyfibrillogenic or insoluble and associated with disease states.

“Aβ amyloidosis” is clinically defined as evidence of Aβ deposition inthe brain or blood vessels of the brain, typically in the form ofamyloid plaques or CAA. Diseases associated with Aβ amyloidosis include,but are not limited to, preclinical Alzheimer's disease, Alzheimer'sdisease (AD), cerebral amyloid angiopathy (CAA), Lewy body dementia, andinclusion body myositis. An “increased risk of developing a diseaseassociated with Aβ amyloidosis” refers to a risk that is elevated overthe expected risk given the subject's age, family history, geneticstatus and other known risk factors.

A “clinical sign of Aβ amyloidosis” refers to a measure of Aβ depositionknown in the art. Clinical signs of Aβ amyloidosis may include, but arenot limited to, Aβ deposition identified by amyloid imaging (e.g. PiBPET, fluorbetapir, or other imaging methods known in the art) or bydecreased cerebrospinal fluid (CSF) Aβ42 or Aβ42/40 ratio. See, forexample, Klunk W E et al. Ann Neurol 55(3) 2004, and Fagan A M et al.Ann Neurol 59(3) 2006, each hereby incorporated by reference in itsentirety. Clinical signs of Aβ amyloidosis may also include measurementsof the metabolism of Aβ, in particular measurements of Aβ42 metabolismalone or in comparison to measurements of the metabolism of other Aβvariants (e.g. Aβ37, Aβ38, Aβ39, Aβ40, and/or total Aβ), as described inU.S. patent Ser. Nos. 14/366,831, 14/523,148 and 14/747,453, each herebyincorporated by reference in its entirety. Additional methods aredescribed in Albert et al. Alzheimer's & Dementia 2007 Vol. 7, pp.170-179; McKhann et al., Alzheimer's & Dementia 2007 Vol. 7, pp.263-269; and Sperling et al. Alzheimer's & Dementia 2007 Vol. 7, pp.280-292, each hereby incorporated by reference in its entirety.Importantly, a subject with clinical signs of Aβ amyloidosis may or maynot have symptoms associated with Aβ deposition. Yet subjects withclinical signs of Aβ amyloidosis are at an increased risk of developinga disease associated with Aβ amyloidosis.

An “Aβ plaque associated symptom” or a “CAA associated symptom” refersto any symptom caused by or associated with the formation of amyloidplaques or CAA, respectively, being composed of regularly orderedfibrillar aggregates called amyloid fibrils. Exemplary Aβ plaqueassociated symptoms may include, but are not limited to, neuronaldegeneration, impaired cognitive function, impaired memory, alteredbehavior, emotional dysregulation, seizures, impaired nervous systemstructure or function, and an increased risk of development or worseningof Alzheimer's disease or CAA. Neuronal degeneration may include achange in structure of a neuron (including molecular changes such asintracellular accumulation of toxic proteins, protein aggregates, etc.and macro level changes such as change in shape or length of axons ordendrites, change in myelin sheath composition, loss of myelin sheath,etc.), a change in function of a neuron, a loss of function of a neuron,death of a neuron, or any combination thereof. Impaired cognitivefunction may include but is not limited to difficulties with memory,attention, concentration, language, abstract thought, creativity,executive function, planning, and organization. Altered behavior mayinclude, but is not limited to, physical or verbal aggression,impulsivity, decreased inhibition, apathy, decreased initiation, changesin personality, abuse of alcohol, tobacco or drugs, and otheraddiction-related behaviors. Emotional dysregulation may include, but isnot limited to, depression, anxiety, mania, irritability, and emotionalincontinence. Seizures may include but are not limited to generalizedtonic-clonic seizures, complex partial seizures, and non-epileptic,psychogenic seizures. Impaired nervous system structure or function mayinclude, but is not limited to, hydrocephalus, Parkinsonism, sleepdisorders, psychosis, impairment of balance and coordination. This mayinclude motor impairments such as monoparesis, hemiparesis,tetraparesis, ataxia, ballismus and tremor. This also may includesensory loss or dysfunction including olfactory, tactile, gustatory,visual and auditory sensation. Furthermore, this may include autonomicnervous system impairments such as bowel and bladder dysfunction, sexualdysfunction, blood pressure and temperature dysregulation. Finally, thismay include hormonal impairments attributable to dysfunction of thehypothalamus and pituitary gland such as deficiencies and dysregulationof growth hormone, thyroid stimulating hormone, lutenizing hormone,follicle stimulating hormone, gonadotropin releasing hormone, prolactin,and numerous other hormones and modulators.

“ApoE” (NP_000032.1, UniProtKB Identifier P02649) is an apolipoproteinexpressed from the APOE gene mapped to chromosome 19 (for example, thenucleotide sequence identified as GenBank Accession Number NM_000041, orNCBI Reference Sequence: NC_000019.10), with three major polymorphicforms: ApoE2 (Cys112, Cys158), ApoE3 (Cys112, Arg158), and ApoE4(Arg112, Arg158). Unless expressly stated otherwise, “ApoE” refers to“human ApoE,” and includes functional fragments. “Recombinant ApoE”refers to ApoE encoded by a nucleic acid that has been introduced into asystem (e.g. a prokaryotic cell, a eukaryotic cell, or a cell-freeexpression system) that supports expression of the nucleic acid and itstranslation into a protein. Methods for producing recombinant proteinsare well-known in the art, and the production of recombinant ApoEdisclosed herein is not limited to a particular system. Those of skillin the art will, however, appreciate that the choice of system caninfluence how “free from lipid” ApoE will be when recombinantlyproduced. As used herein, the term “alipidated ApoE” refers to ApoErecombinantly produced in a prokaryotic cell.

The term “antibody,” as used herein, is used in the broadest sense andencompasses various antibody and antibody-like structures, including butnot limited to full-length monoclonal, polyclonal, and multispecific(e.g., bispecific, trispecific, etc.) antibodies, as well as heavy chainantibodies and antibody fragments provided they exhibit the desiredantigen-binding activity. The domain(s) of an antibody that is involvedin binding an antigen is referred to as a “variable region” or “variabledomain,” and is described in further detail below. A single variabledomain may be sufficient to confer antigen-binding specificity.Preferably, but not necessarily, antibodies useful in the discovery areproduced recombinantly. Antibodies may or may not be glycosylated,though glycosylated antibodies may be preferred. An “isolated” antibodyis one which has been separated from a component of its naturalenvironment. In some embodiments, an antibody is purified to greaterthan 95% or 99% purity as determined by methods known in the art.

In addition to antibodies described herein, it may be possible to designan antibody mimetic or an aptamer using methods known in the art thatfunctions substantially the same as an antibody of the invention. An“antibody mimetic” refers to a polypeptide or a protein that canspecifically bind to an antigen but is not structurally related to anantibody. Antibody mimetics have a mass of about 3 kDa to about 20 kDa.Non-limiting examples of antibody mimetics are affibody molecules,affilins, affimers, alphabodies, anticalins, avimers, DARPins, andmonobodies. Aptamers are a class of small nucleic acid ligands that arecomposed of RNA or single-stranded DNA oligonucleotides and have highspecificity and affinity for their targets. Aptamers interact with andbind to their targets through structural recognition, a process similarto that of an antigen-antibody reaction. Aptamers have a lower molecularweight than antibodies, typically about 8-25 kDa.

The terms “full length antibody” and “intact antibody” may be usedinterchangeably, and refer to an antibody having a structuresubstantially similar to a native antibody structure or having heavychains that contain an Fc region as defined herein. The basic structuralunit of a native antibody comprises a tetramer. Each tetramer iscomposed of two identical pairs of polypeptide chains, each pair havingone “light” chain (about 25 kDa) and one “heavy” chain (about 50-70kDa). Light chains are classified as gamma, mu, alpha, and lambda. Heavychains are classified as gamma, mu, alpha, delta, or epsilon, and definethe antibody's isotype as IgG, IgM, IgA, IgD and IgE, respectively. Theamino-terminal portion of each light and heavy chain includes a variableregion of about 100 to 110 or more amino acid sequences primarilyresponsible for antigen recognition (VL and VH, respectively). Thecarboxy-terminal portion of each chain defines a constant regionprimarily responsible for effector function. Within light and heavychains, the variable and constant regions are joined by a “J” region ofabout 12 or more amino acid sequences, with the heavy chain alsoincluding a “D” region of about 10 more amino acid sequences. Intactantibodies are properly cross-linked via disulfide bonds, as is known inthe art.

The variable domains of the heavy chain and light chain of an antibodygenerally have similar structures, with each domain comprising fourconserved framework regions (FRs) and three hypervariable regions(HVRs). (See, e.g., Kindt et al. Kuby Immunology, 6^(th) ed., W.H.Freeman and Co., page 91 (2007).) A single VH or VL domain may besufficient to confer antigen-binding specificity. Furthermore,antibodies that bind a particular antigen may be isolated using a VH orVL domain from an antibody that binds the antigen to screen a library ofcomplementary VL or VH domains, respectively. See, e.g., Portolano etal., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628(1991).

“Framework region” or “FR” refers to variable domain residues other thanhypervariable region (HVR) residues. The FR of a variable domaingenerally consists of four FR domains: FR1, FR2, FR3, and FR4.Accordingly, the HVR and FR sequences generally appear in the followingsequence: FR1-HVR1-FR2-HVR2-FR3-HVR3-FR4. The FR domains of a heavychain and a light chain may differ, as is known in the art.

The term “hypervariable region” or “HVR” as used herein refers to eachof the regions of a variable domain which are hypervariable in sequence(also commonly referred to as “complementarity determining regions” or“CDR”) and/or form structurally defined loops (“hypervariable loops”)and/or contain the antigen-contacting residues (“antigen contacts”).Generally, antibodies comprise six HVRs: three in the VH (H1, H2, H3),and three in the VL (L1, L2, L3). As used herein, “an HVR derived from avariable region” refers to an HVR that has no more than two amino acidsubstitutions, as compared to the corresponding HVR from the originalvariable region. Exemplary HVRs herein include: (a) hypervariable loopsoccurring at amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3),26-32 (H1), 53-55 (H2), and 96-101 (H3) (Chothia and Lesk, J. Mol. Biol.196:901-917 (1987)); (b) CDRs occurring at amino acid residues 24-34(L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3)(Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed.Public Health Service, National Institutes of Health, Bethesda, Md.(1991)); (c) antigen contacts occurring at amino acid residues 27c-36(L1), 46-55 (L2), 89-96 (L3), 30-35b (H1), 47-58 (H2), and 93-101 (H3)(MacCallum et al. J. Mol. Biol. 262: 732-745 (1996)); and (d)combinations of (a), (b), and/or (c), as defined below for variousantibodies of this disclosure. Unless otherwise indicated, HVR residuesand other residues in the variable domain (e.g., FR residues) arenumbered herein according to Kabat et al., supra.

The term “Fc region” herein is used to define a C-terminal region of animmunoglobulin heavy chain that contains at least a portion of theconstant region. The term includes native sequence Fc regions andvariant Fc regions. In one embodiment, a human IgG heavy chain Fc regionextends from Cys226, or from Pro230, to the carboxyl-terminus of theheavy chain. However, the C-terminal lysine (Lys447) of the Fc regionmay or may not be present. Unless otherwise specified herein, numberingof amino acid residues in the Fc region or constant region is accordingto the EU numbering system, also called the EU index, as described inKabat et al., Sequences of Proteins of Immunological Interest, 5th Ed.Public Health Service, National Institutes of Health, Bethesda, Md.,1991.

A “variant Fc region” comprises an amino acid sequence that can differfrom that of a native Fc region by virtue of one or more amino acidsubstitution(s) and/or by virtue of a modified glycosylation pattern, ascompared to a native Fc region or to the Fc region of a parentpolypeptide. In an example, a variant Fc region can have from about oneto about ten amino acid substitutions, or from about one to about fiveamino acid substitutions in a native sequence Fc region or in the Fcregion of the parent polypeptide. The variant Fc region herein maypossess at least about 80% homology, at least about 90% homology, or atleast about 95% homology with a native sequence Fc region and/or with anFc region of a parent polypeptide.

An “antibody fragment” refers to a molecule other than an intactantibody that comprises a portion of an intact antibody that binds theantigen to which the intact antibody binds. Non-limiting examples ofantibody fragments include but are not limited to Fv, Fab, Fab′,Fab′-SH, F(ab′)₂; single-chain forms of antibodies and higher ordervariants thereof; single-domain antibodies, and multispecific antibodiesformed from antibody fragments.

Single-chain forms of antibodies, and their higher order forms, mayinclude, but are not limited to, single-domain antibodies, single chainvariant fragments (scFvs), divalent scFvs (di-scFvs), trivalent scFvs(tri-scFvs), tetravalent scFvs (tetra-scFvs), diabodies, and triabodiesand tetrabodies. ScFv's are comprised of heavy and light chain variableregions connected by a linker. In most instances, but not all, thelinker may be a peptide. A linker peptide is preferably from about 5 to30 amino acids in length, or from about 10 to 25 amino acids in length.Typically, the linker allows for stabilization of the variable domainswithout interfering with the proper folding and creation of an activebinding site. In preferred embodiments, a linker peptide is rich inglycine, as well as serine or threonine. ScFvs can be used to facilitatephage display or can be used for flow cytometry, immunohistochemistry,or as targeting domains. Methods of making and using scFvs are known inthe art. ScFvs may also be conjugated to a human constant domain (e.g. aheavy constant domain is derived from an IgG domain, such as IgG1, IgG2,IgG3, or IgG4, or a heavy chain constant domain derived from IgA, IgM,or IgE). Diabodies, triabodies, and tetrabodies and higher ordervariants are typically created by varying the length of the linkerpeptide from zero to several amino acids. Alternatively, it is also wellknown in the art that multivalent binding antibody variants can begenerated using self-assembling units linked to the variable domain.

A “single-domain antibody” refers to an antibody fragment consisting ofa single, monomeric variable antibody domain.

Multispecific antibodies include bi-specific antibodies, tri-specific,or antibodies of four or more specificities. Multispecific antibodiesmay be created by combining the heavy and light chains of one antibodywith the heavy and light chains of one or more other antibodies. Thesechains can be covalently linked.

“Monoclonal antibody” refers to an antibody that is derived from asingle copy or clone, including e.g., any eukaryotic, prokaryotic, orphage clone. “Monoclonal antibody” is not limited to antibodies producedthrough hybridoma technology. Monoclonal antibodies can be producedusing hybridoma techniques well known in the art, as well as recombinanttechnologies, phage display technologies, synthetic technologies orcombinations of such technologies and other technologies readily knownin the art. Furthermore, the monoclonal antibody may be labeled with adetectable label, immobilized on a solid phase and/or conjugated with aheterologous compound (e.g., an enzyme or toxin) according to methodsknown in the art.

A “heavy chain antibody” refers to an antibody that consists of twoheavy chains. A heavy chain antibody may be an IgG-like antibody fromcamels, llamas, alpacas, sharks, etc., or an IgNAR from a cartiliaginousfish.

A “humanized antibody” refers to a non-human antibody that has beenmodified to reduce the risk of the non-human antibody eliciting animmune response in humans following administration but retains similarbinding specificity and affinity as the starting non-human antibody. Ahumanized antibody binds to the same or similar epitope as the non-humanantibody. The term “humanized antibody” includes an antibody that iscomposed partially or fully of amino acid sequences derived from a humanantibody germline by altering the sequence of an antibody havingnon-human hypervariable regions (“HVR”). The simplest such alterationmay consist simply of substituting the constant region of a humanantibody for the murine constant region, thus resulting in ahuman/murine chimera which may have sufficiently low immunogenicity tobe acceptable for pharmaceutical use. Preferably, the variable region ofthe antibody is also humanized by techniques that are by now well knownin the art. For example, the framework regions of a variable region canbe substituted by the corresponding human framework regions, whileretaining one, several, or all six non-human HVRs. Some frameworkresidues can be substituted with corresponding residues from a non-humanVL domain or VH domain (e.g., the non-human antibody from which the HVRresidues are derived), e.g., to restore or improve specificity oraffinity of the humanized antibody. Substantially human frameworkregions have at least about 75% homology with a known human frameworksequence (i.e. at least about 75%, at least about 80%, at least about85%, at least about 90%, at least about 95%, or at least about 99%sequence identity). HVRs may also be randomly mutated such that bindingactivity and affinity for the antigen is maintained or enhanced in thecontext of fully human germline framework regions or framework regionsthat are substantially human. As mentioned above, it is sufficient foruse in the methods of this discovery to employ an antibody fragment.Further, as used herein, the term “humanized antibody” refers to anantibody comprising a substantially human framework region, at least oneHVR from a nonhuman antibody, and in which any constant region presentis substantially human. Substantially human constant regions have atleast about 90% with a known human constant sequence (i.e. about 90%,about 95%, or about 99% sequence identity). Hence, all parts of ahumanized antibody, except possibly the HVRs, are substantiallyidentical to corresponding pairs of one or more germline humanimmunoglobulin sequences.

If desired, the design of humanized immunoglobulins may be carried outas follows, or using similar methods familiar to those with skill in theart (for example, see Almagro, et al. Front. Biosci. 2008,13(5):1619-33). A murine antibody variable region is aligned to the mostsimilar human germline sequences (e.g. by using BLAST or similaralgorithm). The CDR residues from the murine antibody sequence aregrafted into the similar human “acceptor” germline. Subsequently, one ormore positions near the CDRs or within the framework (e.g., Vernierpositions) may be reverted to the original murine amino acid in order toachieve a humanized antibody with similar binding affinity to theoriginal murine antibody. Typically, several versions of humanizedantibodies with different reversion mutations are generated andempirically tested for activity. The humanized antibody variant withproperties most similar to the parent murine antibody and the fewestmurine framework reversions is selected as the final humanized antibodycandidate.

II. Anti-ApoE Antibody

Anti-ApoE antibodies disclosed herein can be described or specified interms of the epitope(s) that they recognize or bind. The portion of atarget polypeptide that specifically interacts with the antigen bindingdomain of an antibody is an “epitope.” ApoE can comprise any number ofepitopes, depending on the source of the protein (e.g. mouse, rat,cynomolgus monkey, human, etc.), isoform (e.g. ApoE2, ApoE3, ApoE4),conformational state of the isoform (e.g. fibrillar, aggregated,insoluble, soluble, monomeric, oligomeric, oxidized,post-translationally modified, etc.) and location of the isoform (e.g.intracellular, extracellular, complexed with other proteins or moleculesin particle, in amyloid plaque, etc.). Furthermore, it should be notedthat an “epitope” on ApoE can be a linear epitope or a conformationalepitope, and in both instances can include non-polypeptide elements,e.g., an epitope can include a carbohydrate or lipid side chain. Theterm “affinity” refers to a measure of the strength of the binding of anindividual epitope with an antibody's antigen binding site.

An “anti-ApoE antibody,” as used herein, refers to an isolated antibodythat binds to recombinant human ApoE4 or ApoE4 isolated from human brainwith an affinity constant or affinity of interaction (KD) between about0.1 pM to about 10 μM, preferably about 0.1 pM to about 1 μM, morepreferably about 0.1 pM to about 100 nM. Methods for determining theaffinity of an antibody for an antigen are known in the art, and furtherillustrated in the Examples. Anti-ApoE antibodies useful herein includethose which are suitable for administration to a subject in atherapeutic amount.

Anti-ApoE antibodies disclosed herein can also be described or specifiedin terms of their cross-reactivity. The term “cross-reactivity” refersto the ability of an antibody, specific for one antigen, to react with asecond antigen; a measure of relatedness between two different antigenicsubstances. Thus, an antibody is cross-reactive if it binds to anepitope other than the one that induced its formation. Thecross-reactive epitope generally contains many of the same complementarystructural features as the inducing epitope, and in some cases, canactually fit better than the original. For example, certain antibodieshave some degree of cross-reactivity, in that they bind related, butnon-identical epitopes, e.g., epitopes with at least about 85%, at leastabout 90%, or at least about 95% identity (as calculated using methodsknown in the art) to a reference epitope. An antibody can be said tohave little or no cross-reactivity if it does not bind epitopes withless than about 95%, less than about 90%, or less than about 85%identity to a reference epitope. An antibody can be deemed “highlyspecific” for a certain epitope, if it does not bind any other analog,ortholog, or homolog of that epitope.

The epitope(s) to which anti-ApoE antibodies of this disclosure bind maybe unique to ApoE4, may be common to ApoE4 and another ApoE isoform(e.g. ApoE2 and/or ApoE3), or may be an ApoE4 epitope that is related,but not identical, to an epitope in another isoform. In someembodiments, an anti-ApoE antibody does not preferentially bind toApoE2, ApoE3, or ApoE4. In other embodiments, an anti-ApoE antibodypreferentially binds to ApoE4, or preferentially binds to ApoE3 andApoE4. An antibody that preferentially binds to an ApoE isoform binds tothat isoform more readily than it would a different ApoE isoform. By“preferentially binds,” it is meant that the antibody specifically bindsto an epitope of a first antigen more readily than it would bind toanother epitope of the first antigen or another epitope of a secondantigen. In an example, an antibody can be considered to bind a firstepitope preferentially if it binds the first epitope with an off rate(k(off)) that is less than the antibody's k(off) for the second epitope.In another example, an antibody can be considered to bind a firstepitope preferentially if it binds said first epitope with adissociation constant (KD) that is less than the antibody's KD for thesecond epitope. In another example an antibody can be considered to bindan ApoE isoform preferentially if the binding half maximal concentration(EC₅₀) of the antibody for that isoform is at least about 10-fold,50-fold, or 100-fold less than EC₅₀ for the other isoforms as measuredin an ELISA or similar assay. Alternatively, an antibody can bedescribed as not preferentially binding ApoE2, ApoE3, or ApoE4 if theEC₅₀ for the antibody for each of the isoforms varies by less than10-fold.

Another aspect of isolated, anti-ApoE antibodies of this disclosure isthat they bind to human ApoE in amyloid plaque in unfixed brain tissue.“Unfixed brain tissue” refers to brain tissue that is not fixed withparaformaldehyde or other fixatives. An anti-ApoE antibody is describedas “binding to human ApoE in amyloid plaques in unfixed brain tissue”when the staining pattern is consistent with binding to either Aβ inparenchymal brain amyloid plaques or Aβ in deposits around blood vesselsin the brain in the form of CAA.

Another aspect of isolated, anti-ApoE antibodies of this disclosure isthat they preferentially bind recombinant alipidated human ApoE4 ascompared to ApoE4 derived from human plasma or human ApoE4 derived fromplasma of a transgenic mouse expressing human ApoE4. Briefly, this canbe measured by (a) coating recombinant alipidated human ApoE4 on anELISA plate, (b) incubating an anti-ApoE antibody in the presence ofvarying dilutions of human plasma or plasma from a transgenic mouseexpressing human ApoE4 (e.g., 20-fold to 1000-fold dilutions of plasmacomprising about 1-5 μM ApoE) to produce a pre-incubated antibody-plasmamixture for each dilution, (c) adding the a pre-incubatedantibody-plasma mixtures to the ELISA plate from (a) and allowing themixtures to equilibrate with the coated recombinant alipidated ApoE4 onthe plate, and (d) measuring binding of the anti-ApoE antibody in themixtures to the recombinant alipidated ApoE4 coated on the plate. Anantibody that preferentially binds recombinant alipidated ApoE4 willdemonstrate similar binding to the plate at plasma dilutions betweenabout 1000-fold up to about 20-fold (i.e., loss of binding signal nogreater than 20%, or preferably no greater than 10%). Further detailsare provided in Example 13. In certain embodiments, an isolated,anti-ApoE antibody does not specifically bind to ApoE derived fromplasma. Clearance (CL) is a pharmacokinetic parameter that describes theefficiency of irreversible elimination of a drug from systemiccirculation, expressed as volume of blood/plasma/serum cleared of drugper unit time. An anti-ApoE antibody does not specifically bind to ApoEderived from plasma, for example, when it is administered to an animalexpressing human ApoE and the clearance value of the antibody is lessthan 25 times that of an isotype control antibody (the amountsadministered to the animal being similar for both antibodies). Forexample, see FIG. 37F and Example 19.

Another aspect of isolated, anti-ApoE antibodies of this disclosure isthat they may or may not have a variant Fc region. For example, an Fcregion can be modified to have increased or decreased affinity for an Fcreceptor on a microglial cell and/or an altered glycosylation pattern.

Other aspects of anti-ApoE antibodies of this disclosure are describedmore thoroughly below.

(a) Anti-ApoE Antibodies that Preferentially Bind to ApoE3 and ApoE4:Group I

In another aspect, an anti-ApoE antibody has a heavy chain variableregion comprising SEQ ID NO: 94. In some embodiments, the heavy chainvariable region further comprises SEQ ID NO: 90 and/or SEQ ID NO: 92. Inother embodiments, the heavy chain variable region further comprises SEQID NO: 91 and/or SEQ ID NO: 93. In certain of the above embodiments, theantibody has a light chain variable region comprising SEQ ID NO: 88 orSEQ ID NO: 89. The light chain variable region can further comprise (a)SEQ ID NO: 86 or SEQ ID NO: 87; and/or (b) SEQ ID NO: 30.

In another aspect, an anti-ApoE antibody has a heavy chain variableregion comprising SEQ ID NO: 95. In some embodiments, the heavy chainvariable region further comprises SEQ ID NO: 90 and/or SEQ ID NO: 92. Inother embodiments, the heavy chain variable region further comprises SEQID NO: 91 and/or SEQ ID NO: 93. In certain of the above embodiments, theantibody has a light chain variable region comprising SEQ ID NO: 88 orSEQ ID NO: 89. The light chain variable region can further comprise (a)SEQ ID NO: 86 or SEQ ID NO: 87; and/or (b) SEQ ID NO: 30.

In another aspect, an anti-ApoE antibody is selected from Table A.

TABLE A Group I antibodies Light Chain HVR Heavy Chain HVR Antibody L1L2 L3 H1 H2 H3 1 SEQ ID NO: 86 2 SEQ ID NO: 86 SEQ ID NO: 30 3 SEQ IDNO: 86 SEQ ID NO: 30 SEQ ID NO: 88 4 SEQ ID NO: 30 5 SEQ ID NO: 30 SEQID NO: 88 6 SEQ ID NO: 88 7 SEQ ID NO: 86 SEQ ID NO: 88 8 SEQ ID NO: 909 SEQ ID NO: 90 SEQ ID NO: 92 10 SEQ ID NO: 90 SEQ ID NO: 92 SEQ ID NO:94 11 SEQ ID NO: 92 12 SEQ ID NO: 92 SEQ ID NO: 94 13 SEQ ID NO: 94 14SEQ ID NO: 90 SEQ ID NO: 94 15 SEQ ID NO: 86 SEQ ID NO: 90 16 SEQ ID NO:86 SEQ ID NO: 90 SEQ ID NO: 92 17 SEQ ID NO: 86 SEQ ID NO: 90 SEQ ID NO:92 SEQ ID NO: 94 18 SEQ ID NO: 86 SEQ ID NO: 92 19 SEQ ID NO: 86 SEQ IDNO: 92 SEQ ID NO: 94 20 SEQ ID NO: 86 SEQ ID NO: 94 21 SEQ ID NO: 86 SEQID NO: 90 SEQ ID NO: 94 22 SEQ ID NO: 86 SEQ ID NO: 30 SEQ ID NO: 90 23SEQ ID NO: 86 SEQ ID NO: 30 SEQ ID NO: 90 SEQ ID NO: 92 24 SEQ ID NO: 86SEQ ID NO: 30 SEQ ID NO: 90 SEQ ID NO: 92 SEQ ID NO: 94 25 SEQ ID NO: 86SEQ ID NO: 30 SEQ ID NO: 92 26 SEQ ID NO: 86 SEQ ID NO: 30 SEQ ID NO: 92SEQ ID NO: 94 27 SEQ ID NO: 86 SEQ ID NO: 30 SEQ ID NO: 94 28 SEQ ID NO:86 SEQ ID NO: 30 SEQ ID NO: 90 SEQ ID NO: 94 29 SEQ ID NO: 86 SEQ ID NO:30 SEQ ID NO: 88 SEQ ID NO: 90 30 SEQ ID NO: 86 SEQ ID NO: 30 SEQ ID NO:88 SEQ ID NO: 90 SEQ ID NO: 92 31 SEQ ID NO: 86 SEQ ID NO: 30 SEQ ID NO:88 SEQ ID NO: 90 SEQ ID NO: 92 SEQ ID NO: 94 32 SEQ ID NO: 86 SEQ ID NO:30 SEQ ID NO: 88 SEQ ID NO: 92 33 SEQ ID NO: 86 SEQ ID NO: 30 SEQ ID NO:88 SEQ ID NO: 92 SEQ ID NO: 94 34 SEQ ID NO: 86 SEQ ID NO: 30 SEQ ID NO:88 SEQ ID NO: 90 SEQ ID NO: 94 35 SEQ ID NO: 86 SEQ ID NO: 30 SEQ ID NO:88 SEQ ID NO: 94 36 SEQ ID NO: 30 SEQ ID NO: 90 37 SEQ ID NO: 30 SEQ IDNO: 90 SEQ ID NO: 92 38 SEQ ID NO: 30 SEQ ID NO: 90 SEQ ID NO: 92 SEQ IDNO: 94 39 SEQ ID NO: 30 SEQ ID NO: 92 40 SEQ ID NO: 30 SEQ ID NO: 92 SEQID NO: 94 41 SEQ ID NO: 30 SEQ ID NO: 94 42 SEQ ID NO: 30 SEQ ID NO: 90SEQ ID NO: 94 43 SEQ ID NO: 30 SEQ ID NO: 88 SEQ ID NO: 90 44 SEQ ID NO:30 SEQ ID NO: 88 SEQ ID NO: 90 SEQ ID NO: 92 45 SEQ ID NO: 30 SEQ ID NO:88 SEQ ID NO: 90 SEQ ID NO: 92 SEQ ID NO: 94 46 SEQ ID NO: 30 SEQ ID NO:88 SEQ ID NO: 92 47 SEQ ID NO: 30 SEQ ID NO: 88 SEQ ID NO: 92 SEQ ID NO:94 48 SEQ ID NO: 30 SEQ ID NO: 88 SEQ ID NO: 94 49 SEQ ID NO: 30 SEQ IDNO: 88 SEQ ID NO: 90 SEQ ID NO: 94 50 SEQ ID NO: 88 SEQ ID NO: 90 51 SEQID NO: 88 SEQ ID NO: 90 SEQ ID NO: 92 52 SEQ ID NO: 88 SEQ ID NO: 90 SEQID NO: 92 SEQ ID NO: 94 53 SEQ ID NO: 88 SEQ ID NO: 92 54 SEQ ID NO: 88SEQ ID NO: 92 SEQ ID NO: 94 55 SEQ ID NO: 88 SEQ ID NO: 94 56 SEQ ID NO:88 SEQ ID NO: 90 SEQ ID NO: 94 57 SEQ ID NO: 86 SEQ ID NO: 88 SEQ ID NO:90 58 SEQ ID NO: 86 SEQ ID NO: 88 SEQ ID NO: 90 SEQ ID NO: 92 59 SEQ IDNO: 86 SEQ ID NO: 88 SEQ ID NO: 90 SEQ ID NO: 92 SEQ ID NO: 94 60 SEQ IDNO: 86 SEQ ID NO: 88 SEQ ID NO: 92 61 SEQ ID NO: 86 SEQ ID NO: 88 SEQ IDNO: 92 SEQ ID NO: 94 62 SEQ ID NO: 86 SEQ ID NO: 88 SEQ ID NO: 94 63 SEQID NO: 86 SEQ ID NO: 88 SEQ ID NO: 90 SEQ ID NO: 94 64 SEQ ID NO: 87 65SEQ ID NO: 87 SEQ ID NO: 30 66 SEQ ID NO: 87 SEQ ID NO: 30 SEQ ID NO: 8967 SEQ ID NO: 30 68 SEQ ID NO: 30 SEQ ID NO: 89 69 SEQ ID NO: 89 70 SEQID NO: 87 SEQ ID NO: 89 71 SEQ ID NO: 91 72 SEQ ID NO: 91 SEQ ID NO: 9373 SEQ ID NO: 91 SEQ ID NO: 93 SEQ ID NO: 95 74 SEQ ID NO: 93 75 SEQ IDNO: 93 SEQ ID NO: 95 76 SEQ ID NO: 95 77 SEQ ID NO: 91 SEQ ID NO: 95 78SEQ ID NO: 87 SEQ ID NO: 91 79 SEQ ID NO: 87 SEQ ID NO: 91 SEQ ID NO: 9380 SEQ ID NO: 87 SEQ ID NO: 91 SEQ ID NO: 93 SEQ ID NO: 95 81 SEQ ID NO:87 SEQ ID NO: 93 82 SEQ ID NO: 87 SEQ ID NO: 93 SEQ ID NO: 95 83 SEQ IDNO: 87 SEQ ID NO: 95 84 SEQ ID NO: 87 SEQ ID NO: 91 SEQ ID NO: 95 85 SEQID NO: 87 SEQ ID NO: 30 SEQ ID NO: 91 86 SEQ ID NO: 87 SEQ ID NO: 30 SEQID NO: 91 SEQ ID NO: 93 87 SEQ ID NO: 87 SEQ ID NO: 30 SEQ ID NO: 91 SEQID NO: 93 SEQ ID NO: 95 88 SEQ ID NO: 87 SEQ ID NO: 30 SEQ ID NO: 93 89SEQ ID NO: 87 SEQ ID NO: 30 SEQ ID NO: 93 SEQ ID NO: 95 90 SEQ ID NO: 87SEQ ID NO: 30 SEQ ID NO: 95 91 SEQ ID NO: 87 SEQ ID NO: 30 SEQ ID NO: 91SEQ ID NO: 95 92 SEQ ID NO: 87 SEQ ID NO: 30 SEQ ID NO: 89 SEQ ID NO: 9193 SEQ ID NO: 87 SEQ ID NO: 30 SEQ ID NO: 89 SEQ ID NO: 91 SEQ ID NO: 9394 SEQ ID NO: 87 SEQ ID NO: 30 SEQ ID NO: 89 SEQ ID NO: 91 SEQ ID NO: 93SEQ ID NO: 95 95 SEQ ID NO: 87 SEQ ID NO: 30 SEQ ID NO: 89 SEQ ID NO: 9396 SEQ ID NO: 87 SEQ ID NO: 30 SEQ ID NO: 89 SEQ ID NO: 93 SEQ ID NO: 9597 SEQ ID NO: 87 SEQ ID NO: 30 SEQ ID NO: 89 SEQ ID NO: 95 98 SEQ ID NO:87 SEQ ID NO: 30 SEQ ID NO: 89 SEQ ID NO: 91 SEQ ID NO: 95 99 SEQ ID NO:30 SEQ ID NO: 91 100 SEQ ID NO: 30 SEQ ID NO: 91 SEQ ID NO: 93 101 SEQID NO: 30 SEQ ID NO: 91 SEQ ID NO: 93 SEQ ID NO: 95 102 SEQ ID NO: 30SEQ ID NO: 93 103 SEQ ID NO: 30 SEQ ID NO: 93 SEQ ID NO: 95 104 SEQ IDNO: 30 SEQ ID NO: 95 105 SEQ ID NO: 30 SEQ ID NO: 91 SEQ ID NO: 95 106SEQ ID NO: 30 SEQ ID NO: 89 SEQ ID NO: 91 107 SEQ ID NO: 30 SEQ ID NO:89 SEQ ID NO: 91 SEQ ID NO: 93 108 SEQ ID NO: 30 SEQ ID NO: 89 SEQ IDNO: 91 SEQ ID NO: 93 SEQ ID NO: 95 109 SEQ ID NO: 30 SEQ ID NO: 89 SEQID NO: 93 110 SEQ ID NO: 30 SEQ ID NO: 89 SEQ ID NO: 93 SEQ ID NO: 95111 SEQ ID NO: 30 SEQ ID NO: 89 SEQ ID NO: 95 112 SEQ ID NO: 30 SEQ IDNO: 89 SEQ ID NO: 91 SEQ ID NO: 95 113 SEQ ID NO: 89 SEQ ID NO: 91 114SEQ ID NO: 89 SEQ ID NO: 91 SEQ ID NO: 93 115 SEQ ID NO: 89 SEQ ID NO:91 SEQ ID NO: 93 SEQ ID NO: 95 116 SEQ ID NO: 89 SEQ ID NO: 93 117 SEQID NO: 89 SEQ ID NO: 93 SEQ ID NO: 95 118 SEQ ID NO: 89 SEQ ID NO: 95119 SEQ ID NO: 89 SEQ ID NO: 91 SEQ ID NO: 95 120 SEQ ID NO: 87 SEQ IDNO: 89 SEQ ID NO: 91 121 SEQ ID NO: 87 SEQ ID NO: 89 SEQ ID NO: 91 SEQID NO: 93 122 SEQ ID NO: 87 SEQ ID NO: 89 SEQ ID NO: 91 SEQ ID NO: 93SEQ ID NO: 95 123 SEQ ID NO: 87 SEQ ID NO: 89 SEQ ID NO: 93 124 SEQ IDNO: 87 SEQ ID NO: 89 SEQ ID NO: 93 SEQ ID NO: 95 125 SEQ ID NO: 87 SEQID NO: 89 SEQ ID NO: 95 126 SEQ ID NO: 87 SEQ ID NO: 89 SEQ ID NO: 91SEQ ID NO: 95

In an exemplary embodiment, an anti-ApoE antibody of this groupcomprises a VL that has one or more HVRs derived from SEQ ID NO: 3 or aVH that has one or more HVRs derived from SEQ ID NO: 4. The HVR derivedfrom SEQ ID NO: 3 may be L1, L2, L3, or any combination thereof. Incertain embodiments, the VL may comprise an L1 of SEQ ID NO: 29, an L2of SEQ ID NO: 30, an L3 of SEQ ID NO: 31, or any combination thereof(e.g. antibodies 1-7 in Table B). The HVR derived from SEQ ID NO: 4 maybe H1, H2, H3, or any combination thereof. In certain embodiments, theVH may comprise an H1 of SEQ ID NO: 32, an H2 of SEQ ID NO: 33, an H3 ofSEQ ID NO: 34, or any combination thereof (e.g. antibodies 8-14 in TableB). The antibody comprising one or more HVRs derived from SEQ ID NO: 4may further comprise a light chain variable region (VL) comprising oneor more HVRs derived from SEQ ID NO: 3. The HVR may be L1, L2, L3, orany combination thereof. In a preferred embodiment, the VL may comprisean L1 of SEQ ID NO: 29, an L2 of SEQ ID NO: 30, an L3 of SEQ ID NO: 31,or any combination thereof (e.g. antibodies 15-63 in Table B). Invarious embodiments above, the antibody may be a humanized antibody, orthe antibody may have a VL with 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,or 100% identity to SEQ ID NO: 3 and/or a VH with 90, 91, 92, 93, 94,95, 96, 97, 98, 99 or 100% identity to SEQ ID NO: 4. In each of theabove embodiments, the anti-ApoE antibody may optionally comprise one ormore constant regions, or a portion of a constant region, that issubstantially human (i.e. at least 90%, 95%, or 99% sequence identitywith a known human framework sequence). The present disclosure alsoencompasses the corresponding nucleic acid sequences of SEQ ID NO: 3, 4,29, 30, 31, 32, 33, and 34, which can readily be determined by one ofskill in the art, and may be incorporated into a vector or other largeDNA molecule, such as a chromosome, in order to express an antibody ofthe disclosure.

In another exemplary embodiment, an anti-ApoE antibody of this groupcomprises a VL that has one or more HVRs derived from SEQ ID NO: 17 or aVH that has one or more HVRs derived from SEQ ID NO: 18. The HVR derivedfrom SEQ ID NO: 17 may be L1, L2, L3, or any combination thereof. Incertain embodiments, the VL may comprise an L1 of SEQ ID NO: 63, an L2of SEQ ID NO: 30, an L3 of SEQ ID NO: 64, or any combination thereof(e.g. antibodies 64-70 in Table B). The HVR derived from SEQ ID NO: 18may be H1, H2, H3, or any combination thereof. In certain embodiments,the VH may comprise an H1 of SEQ ID NO: 65, an H2 of SEQ ID NO: 66, anH3 of SEQ ID NO: 67, or any combination thereof (e.g. antibodies 71-77in Table B). The antibody comprising one or more HVRs derived from SEQID NO: 18 may further comprise a light chain variable region (VL)comprising one or more HVRs derived from SEQ ID NO: 17. The HVR may beL1, L2, L3, or any combination thereof. In a preferred embodiment, theVL may comprise an L1 of SEQ ID NO: 63, an L2 of SEQ ID NO: 30, an L3 ofSEQ ID NO: 64, or any combination thereof (e.g. antibodies 78-126 inTable B). In various embodiments above, the antibody may be a humanizedantibody, or the antibody may have a VL with 90, 91, 92, 93, 94, 95, 96,97, 98, 99, or 100% identity to SEQ ID NO: 17 and/or a VH with 90, 91,92, 93, 94, 95, 96, 97, 98, 99 or 100% identity to SEQ ID NO: 18. Ineach of the above embodiments, the anti-ApoE antibody may optionallycomprise one or more constant regions, or a portion of a constantregion, that is substantially human (i.e. at least 90%, 95%, or 99%sequence identity with a known human framework sequence). The presentdisclosure also encompasses the corresponding nucleic acid sequences ofSEQ ID NO: 17, 18, 63, 30, 64, 65, 66, and 67, which can readily bedetermined by one of skill in the art, and may be incorporated into avector or other large DNA molecule, such as a chromosome, in order toexpress an antibody of the disclosure.

In some embodiments, each of the exemplary antibodies described abovemay also contain a variant Fc region, including but not limited to avariant Fc region that is modified to alter the natural interaction withthe microglia FcR.

In further embodiments, an isolated antibody of Group I recognizes anepitope listed in Tables 4-7, for example, as described for theexemplary antibody HJ152 or HJ1514.

TABLE B Exemplary Group I Antibodies Light Chain HVR Heavy Chain HVRAntibody L1 L2 L3 H1 H2 H3 1 SEQ ID NO: 29 2 SEQ ID NO: 29 SEQ ID NO: 303 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 31 4 SEQ ID NO: 30 5 SEQ ID NO:30 SEQ ID NO: 31 6 SEQ ID NO: 31 7 SEQ ID NO: 29 SEQ ID NO: 31 8 SEQ IDNO: 32 9 SEQ ID NO: 32 SEQ ID NO: 33 10 SEQ ID NO: 32 SEQ ID NO: 33 SEQID NO: 34 11 SEQ ID NO: 33 12 SEQ ID NO: 33 SEQ ID NO: 34 13 SEQ ID NO:34 14 SEQ ID NO: 32 SEQ ID NO: 34 15 SEQ ID NO: 29 SEQ ID NO: 32 16 SEQID NO: 29 SEQ ID NO: 32 SEQ ID NO: 33 17 SEQ ID NO: 29 SEQ ID NO: 32 SEQID NO: 33 SEQ ID NO: 34 18 SEQ ID NO: 29 SEQ ID NO: 33 19 SEQ ID NO: 29SEQ ID NO: 33 SEQ ID NO: 34 20 SEQ ID NO: 29 SEQ ID NO: 34 21 SEQ ID NO:29 SEQ ID NO: 32 SEQ ID NO: 34 22 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO:32 23 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 32 SEQ ID NO: 33 24 SEQ IDNO: 29 SEQ ID NO: 30 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 25 SEQ IDNO: 29 SEQ ID NO: 30 SEQ ID NO: 33 26 SEQ ID NO: 29 SEQ ID NO: 30 SEQ IDNO: 33 SEQ ID NO: 34 27 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 34 28 SEQID NO: 29 SEQ ID NO: 30 SEQ ID NO: 32 SEQ ID NO: 34 29 SEQ ID NO: 29 SEQID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 30 SEQ ID NO: 29 SEQ ID NO: 30 SEQID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 31 SEQ ID NO: 29 SEQ ID NO: 30 SEQID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 32 SEQ ID NO: 29 SEQID NO: 30 SEQ ID NO: 31 SEQ ID NO: 33 33 SEQ ID NO: 29 SEQ ID NO: 30 SEQID NO: 31 SEQ ID NO: 33 SEQ ID NO: 34 34 SEQ ID NO: 29 SEQ ID NO: 30 SEQID NO: 31 SEQ ID NO: 32 SEQ ID NO: 34 35 SEQ ID NO: 29 SEQ ID NO: 30 SEQID NO: 31 SEQ ID NO: 34 36 SEQ ID NO: 30 SEQ ID NO: 32 37 SEQ ID NO: 30SEQ ID NO: 32 SEQ ID NO: 33 38 SEQ ID NO: 30 SEQ ID NO: 32 SEQ ID NO: 33SEQ ID NO: 34 39 SEQ ID NO: 30 SEQ ID NO: 33 40 SEQ ID NO: 30 SEQ ID NO:33 SEQ ID NO: 34 41 SEQ ID NO: 30 SEQ ID NO: 34 42 SEQ ID NO: 30 SEQ IDNO: 32 SEQ ID NO: 34 43 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 44 SEQID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 45 SEQ ID NO: 30 SEQID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 46 SEQ ID NO: 30 SEQID NO: 31 SEQ ID NO: 33 47 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 33 SEQID NO: 34 48 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 34 49 SEQ ID NO: 30SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 34 50 SEQ ID NO: 31 SEQ ID NO: 3251 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 52 SEQ ID NO: 31 SEQ ID NO:32 SEQ ID NO: 33 SEQ ID NO: 34 53 SEQ ID NO: 31 SEQ ID NO: 33 54 SEQ IDNO: 31 SEQ ID NO: 33 SEQ ID NO: 34 55 SEQ ID NO: 31 SEQ ID NO: 34 56 SEQID NO: 31 SEQ ID NO: 32 SEQ ID NO: 34 57 SEQ ID NO: 29 SEQ ID NO: 31 SEQID NO: 32 58 SEQ ID NO: 29 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 59SEQ ID NO: 29 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 60SEQ ID NO: 29 SEQ ID NO: 31 SEQ ID NO: 33 61 SEQ ID NO: 29 SEQ ID NO: 31SEQ ID NO: 33 SEQ ID NO: 34 62 SEQ ID NO: 29 SEQ ID NO: 31 SEQ ID NO: 3463 SEQ ID NO: 29 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 34 64 SEQ ID NO:63 65 SEQ ID NO: 63 SEQ ID NO: 30 66 SEQ ID NO: 63 SEQ ID NO: 30 SEQ IDNO: 64 67 SEQ ID NO: 30 68 SEQ ID NO: 30 SEQ ID NO: 64 69 SEQ ID NO: 6470 SEQ ID NO: 63 SEQ ID NO: 64 71 SEQ ID NO: 65 72 SEQ ID NO: 65 SEQ IDNO: 66 73 SEQ ID NO: 65 SEQ ID NO: 66 SEQ ID NO: 67 74 SEQ ID NO: 66 75SEQ ID NO: 66 SEQ ID NO: 67 76 SEQ ID NO: 67 77 SEQ ID NO: 65 SEQ ID NO:67 78 SEQ ID NO: 63 SEQ ID NO: 65 79 SEQ ID NO: 63 SEQ ID NO: 65 SEQ IDNO: 66 80 SEQ ID NO: 63 SEQ ID NO: 65 SEQ ID NO: 66 SEQ ID NO: 67 81 SEQID NO: 63 SEQ ID NO: 66 82 SEQ ID NO: 63 SEQ ID NO: 66 SEQ ID NO: 67 83SEQ ID NO: 63 SEQ ID NO: 67 84 SEQ ID NO: 63 SEQ ID NO: 65 SEQ ID NO: 6785 SEQ ID NO: 63 SEQ ID NO: 30 SEQ ID NO: 65 86 SEQ ID NO: 63 SEQ ID NO:30 SEQ ID NO: 65 SEQ ID NO: 66 87 SEQ ID NO: 63 SEQ ID NO: 30 SEQ ID NO:65 SEQ ID NO: 66 SEQ ID NO: 67 88 SEQ ID NO: 63 SEQ ID NO: 30 SEQ ID NO:66 89 SEQ ID NO: 63 SEQ ID NO: 30 SEQ ID NO: 66 SEQ ID NO: 67 90 SEQ IDNO: 63 SEQ ID NO: 30 SEQ ID NO: 67 91 SEQ ID NO: 63 SEQ ID NO: 30 SEQ IDNO: 65 SEQ ID NO: 67 92 SEQ ID NO: 63 SEQ ID NO: 30 SEQ ID NO: 64 SEQ IDNO: 65 93 SEQ ID NO: 63 SEQ ID NO: 30 SEQ ID NO: 64 SEQ ID NO: 65 SEQ IDNO: 66 94 SEQ ID NO: 63 SEQ ID NO: 30 SEQ ID NO: 64 SEQ ID NO: 65 SEQ IDNO: 66 SEQ ID NO: 67 95 SEQ ID NO: 63 SEQ ID NO: 30 SEQ ID NO: 64 SEQ IDNO: 66 96 SEQ ID NO: 63 SEQ ID NO: 30 SEQ ID NO: 64 SEQ ID NO: 66 SEQ IDNO: 67 97 SEQ ID NO: 63 SEQ ID NO: 30 SEQ ID NO: 64 SEQ ID NO: 67 98 SEQID NO: 63 SEQ ID NO: 30 SEQ ID NO: 64 SEQ ID NO: 65 SEQ ID NO: 67 99 SEQID NO: 30 SEQ ID NO: 65 100 SEQ ID NO: 30 SEQ ID NO: 65 SEQ ID NO: 66101 SEQ ID NO: 30 SEQ ID NO: 65 SEQ ID NO: 66 SEQ ID NO: 67 102 SEQ IDNO: 30 SEQ ID NO: 66 103 SEQ ID NO: 30 SEQ ID NO: 66 SEQ ID NO: 67 104SEQ ID NO: 30 SEQ ID NO: 67 105 SEQ ID NO: 30 SEQ ID NO: 65 SEQ ID NO:67 106 SEQ ID NO: 30 SEQ ID NO: 64 SEQ ID NO: 65 107 SEQ ID NO: 30 SEQID NO: 64 SEQ ID NO: 65 SEQ ID NO: 66 108 SEQ ID NO: 30 SEQ ID NO: 64SEQ ID NO: 65 SEQ ID NO: 66 SEQ ID NO: 67 109 SEQ ID NO: 30 SEQ ID NO:64 SEQ ID NO: 66 110 SEQ ID NO: 30 SEQ ID NO: 64 SEQ ID NO: 66 SEQ IDNO: 67 111 SEQ ID NO: 30 SEQ ID NO: 64 SEQ ID NO: 67 112 SEQ ID NO: 30SEQ ID NO: 64 SEQ ID NO: 65 SEQ ID NO: 67 113 SEQ ID NO: 64 SEQ ID NO:65 114 SEQ ID NO: 64 SEQ ID NO: 65 SEQ ID NO: 66 115 SEQ ID NO: 64 SEQID NO: 65 SEQ ID NO: 66 SEQ ID NO: 67 116 SEQ ID NO: 64 SEQ ID NO: 66117 SEQ ID NO: 64 SEQ ID NO: 66 SEQ ID NO: 67 118 SEQ ID NO: 64 SEQ IDNO: 67 119 SEQ ID NO: 64 SEQ ID NO: 65 SEQ ID NO: 67 120 SEQ ID NO: 63SEQ ID NO: 64 SEQ ID NO: 65 121 SEQ ID NO: 63 SEQ ID NO: 64 SEQ ID NO:65 SEQ ID NO: 66 122 SEQ ID NO: 63 SEQ ID NO: 64 SEQ ID NO: 65 SEQ IDNO: 66 SEQ ID NO: 67 123 SEQ ID NO: 63 SEQ ID NO: 64 SEQ ID NO: 66 124SEQ ID NO: 63 SEQ ID NO: 64 SEQ ID NO: 66 SEQ ID NO: 67 125 SEQ ID NO:63 SEQ ID NO: 64 SEQ ID NO: 67 126 SEQ ID NO: 63 SEQ ID NO: 64 SEQ IDNO: 65 SEQ ID NO: 67(b) Group II: Anti-ApoE Antibodies that Preferentially Bind to ApoE3 andApoE4

In another aspect, an anti-ApoE antibody has a heavy chain variableregion comprising SEQ ID NO: 84. In some embodiments, the heavy chainvariable region further comprises SEQ ID NO: 80 and/or SEQ ID NO: 82. Inother embodiments, the heavy chain variable region further comprises SEQID NO: 81 and/or SEQ ID NO: 83. In certain of the above embodiments, theantibody has a light chain variable region comprising SEQ ID NO: 25. Thelight chain variable region can further comprise (a) SEQ ID NO: 78 orSEQ ID NO: 79; and/or (b) SEQ ID NO: 24.

In another aspect, an anti-ApoE antibody has a heavy chain variableregion comprising SEQ ID NO: 85. In some embodiments, the heavy chainvariable region further comprises SEQ ID NO: 80 and/or SEQ ID NO: 82. Inother embodiments, the heavy chain variable region further comprises SEQID NO: 81 and/or SEQ ID NO: 83. In certain of the above embodiments, theantibody has a light chain variable region comprising SEQ ID NO: 25. Thelight chain variable region can further comprise (a) SEQ ID NO: 78 orSEQ ID NO: 79; and/or (b) SEQ ID NO: 24.

In another aspect, an anti-ApoE antibody is selected from Table C.

TABLE C Group II antibodies Light Chain HVR Heavy Chain HVR Antibody L1L2 L3 H1 H2 H3 1 SEQ ID NO: 78 2 SEQ ID NO: 78 SEQ ID NO: 24 3 SEQ IDNO: 78 SEQ ID NO: 24 SEQ ID NO: 25 4 SEQ ID NO: 24 5 SEQ ID NO: 24 SEQID NO: 25 6 SEQ ID NO: 25 7 SEQ ID NO: 78 SEQ ID NO: 25 8 SEQ ID NO: 809 SEQ ID NO: 80 SEQ ID NO: 82 10 SEQ ID NO: 80 SEQ ID NO: 82 SEQ ID NO:84 11 SEQ ID NO: 82 12 SEQ ID NO: 82 SEQ ID NO: 84 13 SEQ ID NO: 84 14SEQ ID NO: 80 SEQ ID NO: 84 15 SEQ ID NO: 78 SEQ ID NO: 80 16 SEQ ID NO:78 SEQ ID NO: 80 SEQ ID NO: 82 17 SEQ ID NO: 78 SEQ ID NO: 80 SEQ ID NO:82 SEQ ID NO: 84 18 SEQ ID NO: 78 SEQ ID NO: 82 19 SEQ ID NO: 78 SEQ IDNO: 82 SEQ ID NO: 84 20 SEQ ID NO: 78 SEQ ID NO: 84 21 SEQ ID NO: 78 SEQID NO: 80 SEQ ID NO: 84 22 SEQ ID NO: 78 SEQ ID NO: 24 SEQ ID NO: 80 23SEQ ID NO: 78 SEQ ID NO: 24 SEQ ID NO: 80 SEQ ID NO: 82 24 SEQ ID NO: 78SEQ ID NO: 24 SEQ ID NO: 80 SEQ ID NO: 82 SEQ ID NO: 84 25 SEQ ID NO: 78SEQ ID NO: 24 SEQ ID NO: 82 26 SEQ ID NO: 78 SEQ ID NO: 24 SEQ ID NO: 82SEQ ID NO: 84 27 SEQ ID NO: 78 SEQ ID NO: 24 SEQ ID NO: 84 28 SEQ ID NO:78 SEQ ID NO: 24 SEQ ID NO: 80 SEQ ID NO: 84 29 SEQ ID NO: 78 SEQ ID NO:24 SEQ ID NO: 25 SEQ ID NO: 80 30 SEQ ID NO: 78 SEQ ID NO: 24 SEQ ID NO:25 SEQ ID NO: 80 SEQ ID NO: 82 31 SEQ ID NO: 78 SEQ ID NO: 24 SEQ ID NO:25 SEQ ID NO: 80 SEQ ID NO: 82 SEQ ID NO: 84 32 SEQ ID NO: 78 SEQ ID NO:24 SEQ ID NO: 25 SEQ ID NO: 82 33 SEQ ID NO: 78 SEQ ID NO: 24 SEQ ID NO:25 SEQ ID NO: 82 SEQ ID NO: 84 34 SEQ ID NO: 78 SEQ ID NO: 24 SEQ ID NO:25 SEQ ID NO: 80 SEQ ID NO: 84 35 SEQ ID NO: 78 SEQ ID NO: 24 SEQ ID NO:25 SEQ ID NO: 84 36 SEQ ID NO: 24 SEQ ID NO: 80 37 SEQ ID NO: 24 SEQ IDNO: 80 SEQ ID NO: 82 38 SEQ ID NO: 24 SEQ ID NO: 80 SEQ ID NO: 82 SEQ IDNO: 84 39 SEQ ID NO: 24 SEQ ID NO: 82 40 SEQ ID NO: 24 SEQ ID NO: 82 SEQID NO: 84 41 SEQ ID NO: 24 SEQ ID NO: 84 42 SEQ ID NO: 24 SEQ ID NO: 80SEQ ID NO: 84 43 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 80 44 SEQ ID NO:24 SEQ ID NO: 25 SEQ ID NO: 80 SEQ ID NO: 82 45 SEQ ID NO: 24 SEQ ID NO:25 SEQ ID NO: 80 SEQ ID NO: 82 SEQ ID NO: 84 46 SEQ ID NO: 24 SEQ ID NO:25 SEQ ID NO: 82 47 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 82 SEQ ID NO:84 48 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 84 49 SEQ ID NO: 24 SEQ IDNO: 25 SEQ ID NO: 80 SEQ ID NO: 84 50 SEQ ID NO: 25 SEQ ID NO: 80 51 SEQID NO: 25 SEQ ID NO: 80 SEQ ID NO: 82 52 SEQ ID NO: 25 SEQ ID NO: 80 SEQID NO: 82 SEQ ID NO: 84 53 SEQ ID NO: 25 SEQ ID NO: 82 54 SEQ ID NO: 25SEQ ID NO: 82 SEQ ID NO: 84 55 SEQ ID NO: 25 SEQ ID NO: 84 56 SEQ ID NO:25 SEQ ID NO: 80 SEQ ID NO: 84 57 SEQ ID NO: 78 SEQ ID NO: 25 SEQ ID NO:80 58 SEQ ID NO: 78 SEQ ID NO: 25 SEQ ID NO: 80 SEQ ID NO: 82 59 SEQ IDNO: 78 SEQ ID NO: 25 SEQ ID NO: 80 SEQ ID NO: 82 SEQ ID NO: 84 60 SEQ IDNO: 78 SEQ ID NO: 25 SEQ ID NO: 82 61 SEQ ID NO: 78 SEQ ID NO: 25 SEQ IDNO: 82 SEQ ID NO: 84 62 SEQ ID NO: 78 SEQ ID NO: 25 SEQ ID NO: 84 63 SEQID NO: 78 SEQ ID NO: 25 SEQ ID NO: 80 SEQ ID NO: 84 64 SEQ ID NO: 79 65SEQ ID NO: 79 SEQ ID NO: 24 66 SEQ ID NO: 79 SEQ ID NO: 24 SEQ ID NO: 2567 SEQ ID NO: 24 68 SEQ ID NO: 24 SEQ ID NO: 25 69 SEQ ID NO: 25 70 SEQID NO: 79 SEQ ID NO: 25 71 SEQ ID NO: 81 72 SEQ ID NO: 81 SEQ ID NO: 8373 SEQ ID NO: 81 SEQ ID NO: 83 SEQ ID NO: 85 74 SEQ ID NO: 83 75 SEQ IDNO: 83 SEQ ID NO: 85 76 SEQ ID NO: 85 77 SEQ ID NO: 81 SEQ ID NO: 85 78SEQ ID NO: 79 SEQ ID NO: 81 79 SEQ ID NO: 79 SEQ ID NO: 81 SEQ ID NO: 8380 SEQ ID NO: 79 SEQ ID NO: 81 SEQ ID NO: 83 SEQ ID NO: 85 81 SEQ ID NO:79 SEQ ID NO: 83 82 SEQ ID NO: 79 SEQ ID NO: 83 SEQ ID NO: 85 83 SEQ IDNO: 79 SEQ ID NO: 85 84 SEQ ID NO: 79 SEQ ID NO: 81 SEQ ID NO: 85 85 SEQID NO: 79 SEQ ID NO: 24 SEQ ID NO: 81 86 SEQ ID NO: 79 SEQ ID NO: 24 SEQID NO: 81 SEQ ID NO: 83 87 SEQ ID NO: 79 SEQ ID NO: 24 SEQ ID NO: 81 SEQID NO: 83 SEQ ID NO: 85 88 SEQ ID NO: 79 SEQ ID NO: 24 SEQ ID NO: 83 89SEQ ID NO: 79 SEQ ID NO: 24 SEQ ID NO: 83 SEQ ID NO: 85 90 SEQ ID NO: 79SEQ ID NO: 24 SEQ ID NO: 85 91 SEQ ID NO: 79 SEQ ID NO: 24 SEQ ID NO: 81SEQ ID NO: 85 92 SEQ ID NO: 79 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 8193 SEQ ID NO: 79 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 81 SEQ ID NO: 8394 SEQ ID NO: 79 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 81 SEQ ID NO: 83SEQ ID NO: 85 95 SEQ ID NO: 79 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 8396 SEQ ID NO: 79 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 83 SEQ ID NO: 8597 SEQ ID NO: 79 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 85 98 SEQ ID NO:79 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 81 SEQ ID NO: 85 99 SEQ ID NO:24 SEQ ID NO: 81 100 SEQ ID NO: 24 SEQ ID NO: 81 SEQ ID NO: 83 101 SEQID NO: 24 SEQ ID NO: 81 SEQ ID NO: 83 SEQ ID NO: 85 102 SEQ ID NO: 24SEQ ID NO: 83 103 SEQ ID NO: 24 SEQ ID NO: 83 SEQ ID NO: 85 104 SEQ IDNO: 24 SEQ ID NO: 85 105 SEQ ID NO: 24 SEQ ID NO: 81 SEQ ID NO: 85 106SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 81 107 SEQ ID NO: 24 SEQ ID NO:25 SEQ ID NO: 81 SEQ ID NO: 83 108 SEQ ID NO: 24 SEQ ID NO: 25 SEQ IDNO: 81 SEQ ID NO: 83 SEQ ID NO: 85 109 SEQ ID NO: 24 SEQ ID NO: 25 SEQID NO: 83 110 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 83 SEQ ID NO: 85111 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 85 112 SEQ ID NO: 24 SEQ IDNO: 25 SEQ ID NO: 81 SEQ ID NO: 85 113 SEQ ID NO: 25 SEQ ID NO: 81 114SEQ ID NO: 25 SEQ ID NO: 81 SEQ ID NO: 83 115 SEQ ID NO: 25 SEQ ID NO:81 SEQ ID NO: 83 SEQ ID NO: 85 116 SEQ ID NO: 25 SEQ ID NO: 83 117 SEQID NO: 25 SEQ ID NO: 83 SEQ ID NO: 85 118 SEQ ID NO: 25 SEQ ID NO: 85119 SEQ ID NO: 25 SEQ ID NO: 81 SEQ ID NO: 85 120 SEQ ID NO: 79 SEQ IDNO: 25 SEQ ID NO: 81 121 SEQ ID NO: 79 SEQ ID NO: 25 SEQ ID NO: 81 SEQID NO: 83 122 SEQ ID NO: 79 SEQ ID NO: 25 SEQ ID NO: 81 SEQ ID NO: 83SEQ ID NO: 85 123 SEQ ID NO: 79 SEQ ID NO: 25 SEQ ID NO: 83 124 SEQ IDNO: 79 SEQ ID NO: 25 SEQ ID NO: 83 SEQ ID NO: 85 125 SEQ ID NO: 79 SEQID NO: 25 SEQ ID NO: 85 126 SEQ ID NO: 79 SEQ ID NO: 25 SEQ ID NO: 81SEQ ID NO: 85

In an exemplary embodiment, an anti-ApoE antibody of this groupcomprises a VL that has one or more HVRs derived from SEQ ID NO: 9 or aVH that has one or more HVRs derived from SEQ ID NO: 10. The HVR derivedfrom SEQ ID NO: 9 may be L1, L2, L3, or any combination thereof. Incertain embodiments, the VL may comprise an L1 of SEQ ID NO: 47, an L2of SEQ ID NO: 24, an L3 of SEQ ID NO: 25, or any combination thereof(e.g. antibodies 1-7 in Table D). The HVR derived from SEQ ID NO: 10 maybe H1, H2, H3, or any combination thereof. In certain embodiments, theVH may comprise an H1 of SEQ ID NO: 48, an H2 of SEQ ID NO: 49, an H3 ofSEQ ID NO: 50, or any combination thereof (e.g. antibodies 8-14 in TableD). The antibody comprising one or more HVRs derived from SEQ ID NO: 10may further comprise a light chain variable region (VL) comprising oneor more HVRs derived from SEQ ID NO: 9. The HVR may be L1, L2, L3, orany combination thereof. In a preferred embodiment, the VL may comprisean L1 of SEQ ID NO: 47, an L2 of SEQ ID NO: 24, an L3 of SEQ ID NO: 25,or any combination thereof (e.g. antibodies 15-63 in Table D). Invarious embodiments above, the antibody may be a humanized antibody, orthe antibody may have a VL with 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,or 100% identity to SEQ ID NO: 9 and/or a VH with 90, 91, 92, 93, 94,95, 96, 97, 98, 99 or 100% identity to SEQ ID NO: 10. In each of theabove embodiments, the anti-ApoE antibody may optionally comprise one ormore constant regions, or a portion of a constant region, that issubstantially human (i.e. at least 85%, 90%, 95%, or 99% sequenceidentity with a known human framework sequence). The present disclosurealso encompasses the corresponding nucleic acid sequences of SEQ ID NO:9, 10, 47, 24, 25, 48, 49, and 50, which can readily be determined byone of skill in the art, and may be incorporated into a vector or otherlarge DNA molecule, such as a chromosome, in order to express anantibody of the disclosure.

In another exemplary embodiment, an anti-ApoE antibody of this groupcomprises a VL that has one or more HVRs derived from SEQ ID NO: 11 or aVH that has one or more HVRs derived from SEQ ID NO: 12. The HVR derivedfrom SEQ ID NO: 11 may be L1, L2, L3, or any combination thereof. Incertain embodiments, the VL may comprise an L1 of SEQ ID NO: 51, an L2of SEQ ID NO: 24, an L3 of SEQ ID NO: 25, or any combination thereof(e.g. antibodies 64-70 in Table D). The HVR derived from SEQ ID NO: 12may be H1, H2, H3, or any combination thereof. In certain embodiments,the VH may comprise an H1 of SEQ ID NO: 52, an H2 of SEQ ID NO: 53, anH3 of SEQ ID NO: 54, or any combination thereof (e.g. antibodies 71-77in Table D). The antibody comprising one or more HVRs derived from SEQID NO: 12 may further comprise a light chain variable region (VL)comprising one or more HVRs derived from SEQ ID NO: 11. The HVR may beL1, L2, L3, or any combination thereof. In a preferred embodiment, theVL may comprise an L1 of SEQ ID NO: 51, an L2 of SEQ ID NO: 24, an L3 ofSEQ ID NO: 25, or any combination thereof (e.g. antibodies 78-126 inTable D). In various embodiments above, the antibody may be a humanizedantibody, or the antibody may have a VL with 90, 91, 92, 93, 94, 95, 96,97, 98, 99, or 100% identity to SEQ ID NO: 11 and/or a VH with 90, 91,92, 93, 94, 95, 96, 97, 98, 99 or 100% identity to SEQ ID NO: 12. Ineach of the above embodiments, the anti-ApoE antibody may optionallycomprise one or more constant regions, or a portion of a constantregion, that is substantially human (i.e. at least 85%, 90%, 95%, or 99%sequence identity with a known human framework sequence). The presentdisclosure also encompasses the corresponding nucleic acid sequences ofSEQ ID NO: 11, 12, 51, 24, 25, 52, 53, and 54, which can readily bedetermined by one of skill in the art, and may be incorporated into avector or other large DNA molecule, such as a chromosome, in order toexpress an antibody of the disclosure.

In another exemplary embodiment, an anti-ApoE antibody of this groupcomprises a VL that has one or more HVRs derived from SEQ ID NO: 13 or aVH that has one or more HVRs derived from SEQ ID NO: 14. The HVR derivedfrom SEQ ID NO: 13 may be L1, L2, L3, or any combination thereof. Incertain embodiments, the VL may comprise an L1 of SEQ ID NO: 55, an L2of SEQ ID NO: 24, an L3 of SEQ ID NO: 25, or any combination thereof(e.g. antibodies 127-133 in Table D). The HVR derived from SEQ ID NO: 14may be H1, H2, H3, or any combination thereof. In certain embodiments,the VH may comprise an H1 of SEQ ID NO: 56, an H2 of SEQ ID NO: 57, anH3 of SEQ ID NO: 58, or any combination thereof (e.g. antibodies 134-140in Table D). The antibody comprising one or more HVRs derived from SEQID NO: 14 may further comprise a light chain variable region (VL)comprising one or more HVRs derived from SEQ ID NO: 13. The HVR may beL1, L2, L3, or any combination thereof. In a preferred embodiment, theVL may comprise an L1 of SEQ ID NO: 55, an L2 of SEQ ID NO: 24, an L3 ofSEQ ID NO: 25, or any combination thereof (e.g. antibodies 141-189 inTable D). In various embodiments above, the antibody may be a humanizedantibody, or the antibody may have a VL with 90, 91, 92, 93, 94, 95, 96,97, 98, 99, or 100% identity to SEQ ID NO: 13 and/or a VH with 90, 91,92, 93, 94, 95, 96, 97, 98, 99 or 100% identity to SEQ ID NO: 14. Ineach of the above embodiments, the anti-ApoE antibody may optionallycomprise one or more constant regions, or a portion of a constantregion, that is substantially human (i.e. at least 90%, 95%, or 99%sequence identity with a known human framework sequence). The presentdisclosure also encompasses the corresponding nucleic acid sequences ofSEQ ID NO: 13, 14, 55, 24, 25, 56, 57, and 58, which can readily bedetermined by one of skill in the art, and may be incorporated into avector or other large DNA molecule, such as a chromosome, in order toexpress an antibody of the disclosure.

In another exemplary embodiment, an anti-ApoE antibody of this groupcomprises a VL that has one or more HVRs derived from SEQ ID NO: 15 or aVH that has one or more HVRs derived from SEQ ID NO: 16. The HVR derivedfrom SEQ ID NO: 15 may be L1, L2, L3, or any combination thereof. Incertain embodiments, the VL may comprise an L1 of SEQ ID NO: 59, an L2of SEQ ID NO: 24, an L3 of SEQ ID NO: 25, or any combination thereof(e.g. antibodies 190-196 in Table D). The HVR derived from SEQ ID NO: 16may be H1, H2, H3, or any combination thereof. In certain embodiments,the VH may comprise an H1 of SEQ ID NO: 60, an H2 of SEQ ID NO: 61, anH3 of SEQ ID NO: 62, or any combination thereof (e.g. antibodies 197-203in Table D). The antibody comprising one or more HVRs derived from SEQID NO: 16 may further comprise a light chain variable region (VL)comprising one or more HVRs derived from SEQ ID NO: 15. The HVR may beL1, L2, L3, or any combination thereof. In a preferred embodiment, theVL may comprise an L1 of SEQ ID NO: 59, an L2 of SEQ ID NO: 24, an L3 ofSEQ ID NO: 25, or any combination thereof (e.g. antibodies 204-252 inTable D). In various embodiments above, the antibody may be a humanizedantibody, or the antibody may have a VL with 90, 91, 92, 93, 94, 95, 96,97, 98, 99, or 100% identity to SEQ ID NO: 15 and/or a VH with 90, 91,92, 93, 94, 95, 96, 97, 98, 99 or 100% identity to SEQ ID NO: 16. Ineach of the above embodiments, the anti-ApoE antibody may optionallycomprise one or more constant regions, or a portion of a constantregion, that is substantially human (i.e. at least 90%, 95%, or 99%sequence identity with a known human framework sequence). The presentdisclosure also encompasses the corresponding nucleic acid sequences ofSEQ ID NO: 15, 16, 59, 24, 25, 60, 61, and 62, which can readily bedetermined by one of skill in the art, and may be incorporated into avector or other large DNA molecule, such as a chromosome, in order toexpress an antibody of the disclosure.

In another exemplary embodiment, an anti-ApoE antibody of this groupcomprises a VL that has one or more HVRs derived from SEQ ID NO: 19 or aVH that has one or more HVRs derived from SEQ ID NO: 20. The HVR derivedfrom SEQ ID NO: 19 may be L1, L2, L3, or any combination thereof. Incertain embodiments, the VL may comprise an L1 of SEQ ID NO: 68, an L2of SEQ ID NO: 24, an L3 of SEQ ID NO: 25, or any combination thereof(e.g. antibodies 253-259 in Table D). The HVR derived from SEQ ID NO: 20may be H1, H2, H3, or any combination thereof. In certain embodiments,the VH may comprise an H1 of SEQ ID NO: 69, an H2 of SEQ ID NO: 70, anH3 of SEQ ID NO: 71, or any combination thereof (e.g. antibodies 260-266in Table D). The antibody comprising one or more HVRs derived from SEQID NO: 20 may further comprise a light chain variable region (VL)comprising one or more HVRs derived from SEQ ID NO: 19. The HVR may beL1, L2, L3, or any combination thereof. In a preferred embodiment, theVL may comprise an L1 of SEQ ID NO: 68, an L2 of SEQ ID NO: 24, an L3 ofSEQ ID NO: 25, or any combination thereof (e.g. antibodies 267-315 inTable D). In various embodiments above, the antibody may be a humanizedantibody, or the antibody may have a VL with 90, 91, 92, 93, 94, 95, 96,97, 98, 99, or 100% identity to SEQ ID NO: 19 and/or a VH with 90, 91,92, 93, 94, 95, 96, 97, 98, 99 or 100% identity to SEQ ID NO: 20. Ineach of the above embodiments, the anti-ApoE antibody may optionallycomprise one or more constant regions, or a portion of a constantregion, that is substantially human (i.e. at least 90%, 95%, or 99%sequence identity with a known human framework sequence). The presentdisclosure also encompasses the corresponding nucleic acid sequences ofSEQ ID NO: 19, 20, 68, 24, 25, 69, 70, and 71, which can readily bedetermined by one of skill in the art, and may be incorporated into avector or other large DNA molecule, such as a chromosome, in order toexpress an antibody of the disclosure.

In some embodiments, each of the exemplary antibodies described abovemay also contain a variant Fc region, including but not limited to avariant Fc region that is modified to alter the natural interaction withthe microglia FcR.

In further embodiments, an isolated antibody of Group II recognizes anepitope listed in Tables 4-7, for example, as described for theexemplary antibody HJ155, HJ156, HJ159, HJ1513, or HJ1518.

TABLE D Exemplary Group II Antibodies Light Chain HVR Heavy Chain HVRAntibody L1 L2 L3 H1 H2 H3 1 SEQ ID NO: 47 2 SEQ ID NO: 47 SEQ ID NO: 243 SEQ ID NO: 47 SEQ ID NO: 24 SEQ ID NO: 25 4 SEQ ID NO: 24 5 SEQ ID NO:24 SEQ ID NO: 25 6 SEQ ID NO: 25 7 SEQ ID NO: 47 SEQ ID NO: 25 8 SEQ IDNO: 48 9 SEQ ID NO: 48 SEQ ID NO: 49 10 SEQ ID NO: 48 SEQ ID NO: 49 SEQID NO: 50 11 SEQ ID NO: 49 12 SEQ ID NO: 49 SEQ ID NO: 50 13 SEQ ID NO:50 14 SEQ ID NO: 48 SEQ ID NO: 50 15 SEQ ID NO: 47 SEQ ID NO: 48 16 SEQID NO: 47 SEQ ID NO: 48 SEQ ID NO: 49 17 SEQ ID NO: 47 SEQ ID NO: 48 SEQID NO: 49 SEQ ID NO: 50 18 SEQ ID NO: 47 SEQ ID NO: 49 19 SEQ ID NO: 47SEQ ID NO: 49 SEQ ID NO: 50 20 SEQ ID NO: 47 SEQ ID NO: 50 21 SEQ ID NO:47 SEQ ID NO: 48 SEQ ID NO: 50 22 SEQ ID NO: 47 SEQ ID NO: 24 SEQ ID NO:48 23 SEQ ID NO: 47 SEQ ID NO: 24 SEQ ID NO: 48 SEQ ID NO: 49 24 SEQ IDNO: 47 SEQ ID NO: 24 SEQ ID NO: 48 SEQ ID NO: 49 SEQ ID NO: 50 25 SEQ IDNO: 47 SEQ ID NO: 24 SEQ ID NO: 49 26 SEQ ID NO: 47 SEQ ID NO: 24 SEQ IDNO: 49 SEQ ID NO: 50 27 SEQ ID NO: 47 SEQ ID NO: 24 SEQ ID NO: 50 28 SEQID NO: 47 SEQ ID NO: 24 SEQ ID NO: 48 SEQ ID NO: 50 29 SEQ ID NO: 47 SEQID NO: 24 SEQ ID NO: 25 SEQ ID NO: 48 30 SEQ ID NO: 47 SEQ ID NO: 24 SEQID NO: 25 SEQ ID NO: 48 SEQ ID NO: 49 31 SEQ ID NO: 47 SEQ ID NO: 24 SEQID NO: 25 SEQ ID NO: 48 SEQ ID NO: 49 SEQ ID NO: 50 32 SEQ ID NO: 47 SEQID NO: 24 SEQ ID NO: 25 SEQ ID NO: 49 33 SEQ ID NO: 47 SEQ ID NO: 24 SEQID NO: 25 SEQ ID NO: 49 SEQ ID NO: 50 34 SEQ ID NO: 47 SEQ ID NO: 24 SEQID NO: 25 SEQ ID NO: 48 SEQ ID NO: 50 35 SEQ ID NO: 47 SEQ ID NO: 24 SEQID NO: 25 SEQ ID NO: 50 36 SEQ ID NO: 24 SEQ ID NO: 48 37 SEQ ID NO: 24SEQ ID NO: 48 SEQ ID NO: 49 38 SEQ ID NO: 24 SEQ ID NO: 48 SEQ ID NO: 49SEQ ID NO: 50 39 SEQ ID NO: 24 SEQ ID NO: 49 40 SEQ ID NO: 24 SEQ ID NO:49 SEQ ID NO: 50 41 SEQ ID NO: 24 SEQ ID NO: 50 42 SEQ ID NO: 24 SEQ IDNO: 48 SEQ ID NO: 50 43 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 48 44 SEQID NO: 24 SEQ ID NO: 25 SEQ ID NO: 48 SEQ ID NO: 49 45 SEQ ID NO: 24 SEQID NO: 25 SEQ ID NO: 48 SEQ ID NO: 49 SEQ ID NO: 50 46 SEQ ID NO: 24 SEQID NO: 25 SEQ ID NO: 49 47 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 49 SEQID NO: 50 48 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 50 49 SEQ ID NO: 24SEQ ID NO: 25 SEQ ID NO: 48 SEQ ID NO: 50 50 SEQ ID NO: 25 SEQ ID NO: 4851 SEQ ID NO: 25 SEQ ID NO: 48 SEQ ID NO: 49 52 SEQ ID NO: 25 SEQ ID NO:48 SEQ ID NO: 49 SEQ ID NO: 50 53 SEQ ID NO: 25 SEQ ID NO: 49 54 SEQ IDNO: 25 SEQ ID NO: 49 SEQ ID NO: 50 55 SEQ ID NO: 25 SEQ ID NO: 50 56 SEQID NO: 25 SEQ ID NO: 48 SEQ ID NO: 50 57 SEQ ID NO: 47 SEQ ID NO: 25 SEQID NO: 48 58 SEQ ID NO: 47 SEQ ID NO: 25 SEQ ID NO: 48 SEQ ID NO: 49 59SEQ ID NO: 47 SEQ ID NO: 25 SEQ ID NO: 48 SEQ ID NO: 49 SEQ ID NO: 50 60SEQ ID NO: 47 SEQ ID NO: 25 SEQ ID NO: 49 61 SEQ ID NO: 47 SEQ ID NO: 25SEQ ID NO: 49 SEQ ID NO: 50 62 SEQ ID NO: 47 SEQ ID NO: 25 SEQ ID NO: 5063 SEQ ID NO: 47 SEQ ID NO: 25 SEQ ID NO: 48 SEQ ID NO: 50 64 SEQ ID NO:51 65 SEQ ID NO: 51 SEQ ID NO: 24 66 SEQ ID NO: 51 SEQ ID NO: 24 SEQ IDNO: 25 67 SEQ ID NO: 24 68 SEQ ID NO: 24 SEQ ID NO: 25 69 SEQ ID NO: 2570 SEQ ID NO: 51 SEQ ID NO: 25 71 SEQ ID NO: 52 72 SEQ ID NO: 52 SEQ IDNO: 53 73 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 74 SEQ ID NO: 53 75SEQ ID NO: 53 SEQ ID NO: 54 76 SEQ ID NO: 54 77 SEQ ID NO: 52 SEQ ID NO:54 78 SEQ ID NO: 51 SEQ ID NO: 52 79 SEQ ID NO: 51 SEQ ID NO: 52 SEQ IDNO: 53 80 SEQ ID NO: 51 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 81 SEQID NO: 51 SEQ ID NO: 53 82 SEQ ID NO: 51 SEQ ID NO: 53 SEQ ID NO: 54 83SEQ ID NO: 51 SEQ ID NO: 54 84 SEQ ID NO: 51 SEQ ID NO: 52 SEQ ID NO: 5485 SEQ ID NO: 51 SEQ ID NO: 24 SEQ ID NO: 52 86 SEQ ID NO: 51 SEQ ID NO:24 SEQ ID NO: 52 SEQ ID NO: 53 87 SEQ ID NO: 51 SEQ ID NO: 24 SEQ ID NO:52 SEQ ID NO: 53 SEQ ID NO: 54 88 SEQ ID NO: 51 SEQ ID NO: 24 SEQ ID NO:53 89 SEQ ID NO: 51 SEQ ID NO: 24 SEQ ID NO: 53 SEQ ID NO: 54 90 SEQ IDNO: 51 SEQ ID NO: 24 SEQ ID NO: 54 91 SEQ ID NO: 51 SEQ ID NO: 24 SEQ IDNO: 52 SEQ ID NO: 54 92 SEQ ID NO: 51 SEQ ID NO: 24 SEQ ID NO: 25 SEQ IDNO: 52 93 SEQ ID NO: 51 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 52 SEQ IDNO: 53 94 SEQ ID NO: 51 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 52 SEQ IDNO: 53 SEQ ID NO: 54 95 SEQ ID NO: 51 SEQ ID NO: 24 SEQ ID NO: 25 SEQ IDNO: 53 96 SEQ ID NO: 51 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 53 SEQ IDNO: 54 97 SEQ ID NO: 51 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 54 98 SEQID NO: 51 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 52 SEQ ID NO: 54 99 SEQID NO: 24 SEQ ID NO: 52 100 SEQ ID NO: 24 SEQ ID NO: 52 SEQ ID NO: 53101 SEQ ID NO: 24 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 102 SEQ IDNO: 24 SEQ ID NO: 53 103 SEQ ID NO: 24 SEQ ID NO: 53 SEQ ID NO: 54 104SEQ ID NO: 24 SEQ ID NO: 54 105 SEQ ID NO: 24 SEQ ID NO: 52 SEQ ID NO:54 106 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 52 107 SEQ ID NO: 24 SEQID NO: 25 SEQ ID NO: 52 SEQ ID NO: 53 108 SEQ ID NO: 24 SEQ ID NO: 25SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 109 SEQ ID NO: 24 SEQ ID NO:25 SEQ ID NO: 53 110 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 53 SEQ IDNO: 54 111 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 54 112 SEQ ID NO: 24SEQ ID NO: 25 SEQ ID NO: 52 SEQ ID NO: 54 113 SEQ ID NO: 25 SEQ ID NO:52 114 SEQ ID NO: 25 SEQ ID NO: 52 SEQ ID NO: 53 115 SEQ ID NO: 25 SEQID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 116 SEQ ID NO: 25 SEQ ID NO: 53117 SEQ ID NO: 25 SEQ ID NO: 53 SEQ ID NO: 54 118 SEQ ID NO: 25 SEQ IDNO: 54 119 SEQ ID NO: 25 SEQ ID NO: 52 SEQ ID NO: 54 120 SEQ ID NO: 51SEQ ID NO: 25 SEQ ID NO: 52 121 SEQ ID NO: 51 SEQ ID NO: 25 SEQ ID NO:52 SEQ ID NO: 53 122 SEQ ID NO: 51 SEQ ID NO: 25 SEQ ID NO: 52 SEQ IDNO: 53 SEQ ID NO: 54 123 SEQ ID NO: 51 SEQ ID NO: 25 SEQ ID NO: 53 124SEQ ID NO: 51 SEQ ID NO: 25 SEQ ID NO: 53 SEQ ID NO: 54 125 SEQ ID NO:51 SEQ ID NO: 25 SEQ ID NO: 54 126 SEQ ID NO: 51 SEQ ID NO: 25 SEQ IDNO: 52 SEQ ID NO: 54 127 SEQ ID NO: 55 128 SEQ ID NO: 55 SEQ ID NO: 24219 SEQ ID NO: 55 SEQ ID NO: 24 SEQ ID NO: 25 130 SEQ ID NO: 24 131 SEQID NO: 24 SEQ ID NO: 25 132 SEQ ID NO: 25 133 SEQ ID NO: 55 SEQ ID NO:25 134 SEQ ID NO: 56 135 SEQ ID NO: 56 SEQ ID NO: 57 136 SEQ ID NO: 56SEQ ID NO: 57 SEQ ID NO: 58 137 SEQ ID NO: 57 138 SEQ ID NO: 57 SEQ IDNO: 58 139 SEQ ID NO: 58 140 SEQ ID NO: 56 SEQ ID NO: 58 141 SEQ ID NO:55 SEQ ID NO: 56 142 SEQ ID NO: 55 SEQ ID NO: 56 SEQ ID NO: 57 143 SEQID NO: 55 SEQ ID NO: 56 SEQ ID NO: 57 SEQ ID NO: 58 144 SEQ ID NO: 55SEQ ID NO: 57 145 SEQ ID NO: 55 SEQ ID NO: 57 SEQ ID NO: 58 146 SEQ IDNO: 55 SEQ ID NO: 58 147 SEQ ID NO: 55 SEQ ID NO: 56 SEQ ID NO: 58 148SEQ ID NO: 55 SEQ ID NO: 24 SEQ ID NO: 56 149 SEQ ID NO: 55 SEQ ID NO:24 SEQ ID NO: 56 SEQ ID NO: 57 150 SEQ ID NO: 55 SEQ ID NO: 24 SEQ IDNO: 56 SEQ ID NO: 57 SEQ ID NO: 58 151 SEQ ID NO: 55 SEQ ID NO: 24 SEQID NO: 57 152 SEQ ID NO: 55 SEQ ID NO: 24 SEQ ID NO: 57 SEQ ID NO: 58153 SEQ ID NO: 55 SEQ ID NO: 24 SEQ ID NO: 58 154 SEQ ID NO: 55 SEQ IDNO: 24 SEQ ID NO: 56 SEQ ID NO: 58 155 SEQ ID NO: 55 SEQ ID NO: 24 SEQID NO: 25 SEQ ID NO: 56 156 SEQ ID NO: 55 SEQ ID NO: 24 SEQ ID NO: 25SEQ ID NO: 56 SEQ ID NO: 57 157 SEQ ID NO: 55 SEQ ID NO: 24 SEQ ID NO:25 SEQ ID NO: 56 SEQ ID NO: 57 SEQ ID NO: 58 158 SEQ ID NO: 55 SEQ IDNO: 24 SEQ ID NO: 25 SEQ ID NO: 57 159 SEQ ID NO: 55 SEQ ID NO: 24 SEQID NO: 25 SEQ ID NO: 57 SEQ ID NO: 58 160 SEQ ID NO: 55 SEQ ID NO: 24SEQ ID NO: 25 SEQ ID NO: 56 SEQ ID NO: 58 161 SEQ ID NO: 55 SEQ ID NO:24 SEQ ID NO: 25 SEQ ID NO: 58 162 SEQ ID NO: 24 SEQ ID NO: 56 163 SEQID NO: 24 SEQ ID NO: 56 SEQ ID NO: 57 164 SEQ ID NO: 24 SEQ ID NO: 56SEQ ID NO: 57 SEQ ID NO: 58 165 SEQ ID NO: 24 SEQ ID NO: 57 166 SEQ IDNO: 24 SEQ ID NO: 57 SEQ ID NO: 58 167 SEQ ID NO: 24 SEQ ID NO: 58 168SEQ ID NO: 24 SEQ ID NO: 56 SEQ ID NO: 58 169 SEQ ID NO: 24 SEQ ID NO:25 SEQ ID NO: 56 170 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 56 SEQ IDNO: 57 171 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 56 SEQ ID NO: 57 SEQID NO: 58 172 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 57 173 SEQ ID NO:24 SEQ ID NO: 25 SEQ ID NO: 57 SEQ ID NO: 58 174 SEQ ID NO: 24 SEQ IDNO: 25 SEQ ID NO: 58 175 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 56 SEQID NO: 58 176 SEQ ID NO: 25 SEQ ID NO: 56 177 SEQ ID NO: 25 SEQ ID NO:56 SEQ ID NO: 57 178 SEQ ID NO: 25 SEQ ID NO: 56 SEQ ID NO: 57 SEQ IDNO: 58 179 SEQ ID NO: 25 SEQ ID NO: 57 180 SEQ ID NO: 25 SEQ ID NO: 57SEQ ID NO: 58 181 SEQ ID NO: 25 SEQ ID NO: 58 182 SEQ ID NO: 25 SEQ IDNO: 56 SEQ ID NO: 58 183 SEQ ID NO: 55 SEQ ID NO: 25 SEQ ID NO: 56 184SEQ ID NO: 55 SEQ ID NO: 25 SEQ ID NO: 56 SEQ ID NO: 57 185 SEQ ID NO:55 SEQ ID NO: 25 SEQ ID NO: 56 SEQ ID NO: 57 SEQ ID NO: 58 186 SEQ IDNO: 55 SEQ ID NO: 25 SEQ ID NO: 57 187 SEQ ID NO: 55 SEQ ID NO: 25 SEQID NO: 57 SEQ ID NO: 58 188 SEQ ID NO: 55 SEQ ID NO: 25 SEQ ID NO: 58189 SEQ ID NO: 55 SEQ ID NO: 25 SEQ ID NO: 56 SEQ ID NO: 58 190 SEQ IDNO: 59 191 SEQ ID NO: 59 SEQ ID NO: 24 192 SEQ ID NO: 59 SEQ ID NO: 24SEQ ID NO: 25 193 SEQ ID NO: 24 194 SEQ ID NO: 24 SEQ ID NO: 25 195 SEQID NO: 25 196 SEQ ID NO: 59 SEQ ID NO: 25 197 SEQ ID NO: 60 198 SEQ IDNO: 60 SEQ ID NO: 61 199 SEQ ID NO: 60 SEQ ID NO: 61 SEQ ID NO: 62 200SEQ ID NO: 61 201 SEQ ID NO: 61 SEQ ID NO: 62 202 SEQ ID NO: 62 203 SEQID NO: 60 SEQ ID NO: 62 204 SEQ ID NO: 59 SEQ ID NO: 60 205 SEQ ID NO:59 SEQ ID NO: 60 SEQ ID NO: 61 206 SEQ ID NO: 59 SEQ ID NO: 60 SEQ IDNO: 61 SEQ ID NO: 62 207 SEQ ID NO: 59 SEQ ID NO: 61 208 SEQ ID NO: 59SEQ ID NO: 61 SEQ ID NO: 62 209 SEQ ID NO: 59 SEQ ID NO: 62 210 SEQ IDNO: 59 SEQ ID NO: 60 SEQ ID NO: 62 211 SEQ ID NO: 59 SEQ ID NO: 24 SEQID NO: 60 212 SEQ ID NO: 59 SEQ ID NO: 24 SEQ ID NO: 60 SEQ ID NO: 61213 SEQ ID NO: 59 SEQ ID NO: 24 SEQ ID NO: 60 SEQ ID NO: 61 SEQ ID NO:62 214 SEQ ID NO: 59 SEQ ID NO: 24 SEQ ID NO: 61 215 SEQ ID NO: 59 SEQID NO: 24 SEQ ID NO: 61 SEQ ID NO: 62 216 SEQ ID NO: 59 SEQ ID NO: 24SEQ ID NO: 62 217 SEQ ID NO: 59 SEQ ID NO: 24 SEQ ID NO: 60 SEQ ID NO:62 218 SEQ ID NO: 59 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 60 219 SEQID NO: 59 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 60 SEQ ID NO: 61 220SEQ ID NO: 59 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 60 SEQ ID NO: 61SEQ ID NO: 62 221 SEQ ID NO: 59 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO:61 222 SEQ ID NO: 59 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 61 SEQ IDNO: 62 223 SEQ ID NO: 59 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 60 SEQID NO: 62 224 SEQ ID NO: 59 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 62225 SEQ ID NO: 24 SEQ ID NO: 60 226 SEQ ID NO: 24 SEQ ID NO: 60 SEQ IDNO: 61 227 SEQ ID NO: 24 SEQ ID NO: 60 SEQ ID NO: 61 SEQ ID NO: 62 228SEQ ID NO: 24 SEQ ID NO: 61 229 SEQ ID NO: 24 SEQ ID NO: 61 SEQ ID NO:62 230 SEQ ID NO: 24 SEQ ID NO: 62 231 SEQ ID NO: 24 SEQ ID NO: 60 SEQID NO: 62 232 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 60 233 SEQ ID NO:24 SEQ ID NO: 25 SEQ ID NO: 60 SEQ ID NO: 61 234 SEQ ID NO: 24 SEQ IDNO: 25 SEQ ID NO: 60 SEQ ID NO: 61 SEQ ID NO: 62 235 SEQ ID NO: 24 SEQID NO: 25 SEQ ID NO: 61 236 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 61SEQ ID NO: 62 237 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 62 238 SEQ IDNO: 24 SEQ ID NO: 25 SEQ ID NO: 60 SEQ ID NO: 62 239 SEQ ID NO: 25 SEQID NO: 60 240 SEQ ID NO: 25 SEQ ID NO: 60 SEQ ID NO: 61 240 SEQ ID NO:25 SEQ ID NO: 60 SEQ ID NO: 61 SEQ ID NO: 62 242 SEQ ID NO: 25 SEQ IDNO: 61 423 SEQ ID NO: 25 SEQ ID NO: 61 SEQ ID NO: 62 244 SEQ ID NO: 25SEQ ID NO: 62 245 SEQ ID NO: 25 SEQ ID NO: 60 SEQ ID NO: 62 246 SEQ IDNO: 59 SEQ ID NO: 25 SEQ ID NO: 60 247 SEQ ID NO: 59 SEQ ID NO: 25 SEQID NO: 60 SEQ ID NO: 61 248 SEQ ID NO: 59 SEQ ID NO: 25 SEQ ID NO: 60SEQ ID NO: 61 SEQ ID NO: 62 249 SEQ ID NO: 59 SEQ ID NO: 25 SEQ ID NO:61 250 SEQ ID NO: 59 SEQ ID NO: 25 SEQ ID NO: 61 SEQ ID NO: 62 251 SEQID NO: 59 SEQ ID NO: 25 SEQ ID NO: 62 252 SEQ ID NO: 59 SEQ ID NO: 25SEQ ID NO: 60 SEQ ID NO: 62 253 SEQ ID NO: 68 2454 SEQ ID NO: 68 SEQ IDNO: 24 255 SEQ ID NO: 68 SEQ ID NO: 24 SEQ ID NO: 25 256 SEQ ID NO: 24257 SEQ ID NO: 24 SEQ ID NO: 25 258 SEQ ID NO: 25 259 SEQ ID NO: 68 SEQID NO: 25 260 SEQ ID NO: 69 261 SEQ ID NO: 69 SEQ ID NO: 70 262 SEQ IDNO: 69 SEQ ID NO: 70 SEQ ID NO: 71 263 SEQ ID NO: 70 264 SEQ ID NO: 70SEQ ID NO: 71 265 SEQ ID NO: 71 266 SEQ ID NO: 69 SEQ ID NO: 71 267 SEQID NO: 68 SEQ ID NO: 69 268 SEQ ID NO: 68 SEQ ID NO: 69 SEQ ID NO: 70269 SEQ ID NO: 68 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 270 SEQ IDNO: 68 SEQ ID NO: 70 271 SEQ ID NO: 68 SEQ ID NO: 70 SEQ ID NO: 71 272SEQ ID NO: 68 SEQ ID NO: 71 273 SEQ ID NO: 68 SEQ ID NO: 69 SEQ ID NO:71 274 SEQ ID NO: 68 SEQ ID NO: 24 SEQ ID NO: 69 275 SEQ ID NO: 68 SEQID NO: 24 SEQ ID NO: 69 SEQ ID NO: 70 276 SEQ ID NO: 68 SEQ ID NO: 24SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 277 SEQ ID NO: 68 SEQ ID NO:24 SEQ ID NO: 70 278 SEQ ID NO: 68 SEQ ID NO: 24 SEQ ID NO: 70 SEQ IDNO: 71 279 SEQ ID NO: 68 SEQ ID NO: 24 SEQ ID NO: 71 280 SEQ ID NO: 68SEQ ID NO: 24 SEQ ID NO: 69 SEQ ID NO: 71 281 SEQ ID NO: 68 SEQ ID NO:24 SEQ ID NO: 25 SEQ ID NO: 69 282 SEQ ID NO: 68 SEQ ID NO: 24 SEQ IDNO: 25 SEQ ID NO: 69 SEQ ID NO: 70 283 SEQ ID NO: 68 SEQ ID NO: 24 SEQID NO: 25 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 284 SEQ ID NO: 68SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 70 285 SEQ ID NO: 68 SEQ ID NO:24 SEQ ID NO: 25 SEQ ID NO: 70 SEQ ID NO: 71 286 SEQ ID NO: 68 SEQ IDNO: 24 SEQ ID NO: 25 SEQ ID NO: 69 SEQ ID NO: 71 287 SEQ ID NO: 68 SEQID NO: 24 SEQ ID NO: 25 SEQ ID NO: 71 288 SEQ ID NO: 24 SEQ ID NO: 69289 SEQ ID NO: 24 SEQ ID NO: 69 SEQ ID NO: 70 290 SEQ ID NO: 24 SEQ IDNO: 69 SEQ ID NO: 70 SEQ ID NO: 71 291 SEQ ID NO: 24 SEQ ID NO: 70 292SEQ ID NO: 24 SEQ ID NO: 70 SEQ ID NO: 71 293 SEQ ID NO: 24 SEQ ID NO:71 294 SEQ ID NO: 24 SEQ ID NO: 69 SEQ ID NO: 71 295 SEQ ID NO: 24 SEQID NO: 25 SEQ ID NO: 69 296 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 69SEQ ID NO: 70 297 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 69 SEQ ID NO:70 SEQ ID NO: 71 298 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 70 299 SEQID NO: 24 SEQ ID NO: 25 SEQ ID NO: 70 SEQ ID NO: 71 300 SEQ ID NO: 24SEQ ID NO: 25 SEQ ID NO: 71 301 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO:69 SEQ ID NO: 71 302 SEQ ID NO: 25 SEQ ID NO: 69 303 SEQ ID NO: 25 SEQID NO: 69 SEQ ID NO: 70 304 SEQ ID NO: 25 SEQ ID NO: 69 SEQ ID NO: 70SEQ ID NO: 71 305 SEQ ID NO: 25 SEQ ID NO: 70 306 SEQ ID NO: 25 SEQ IDNO: 70 SEQ ID NO: 71 307 SEQ ID NO: 25 SEQ ID NO: 71 308 SEQ ID NO: 25SEQ ID NO: 69 SEQ ID NO: 71 309 SEQ ID NO: 68 SEQ ID NO: 25 SEQ ID NO:69 310 SEQ ID NO: 68 SEQ ID NO: 25 SEQ ID NO: 69 SEQ ID NO: 70 311 SEQID NO: 68 SEQ ID NO: 25 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 312SEQ ID NO: 68 SEQ ID NO: 25 SEQ ID NO: 70 313 SEQ ID NO: 68 SEQ ID NO:25 SEQ ID NO: 70 SEQ ID NO: 71 314 SEQ ID NO: 68 SEQ ID NO: 25 SEQ IDNO: 71 315 SEQ ID NO: 68 SEQ ID NO: 25 SEQ ID NO: 69 SEQ ID NO: 71(c) Group III: Anti-ApoE Antibodies that do not Preferentially BindApoE4, ApoE3 or ApoE2

In another aspect, an anti-ApoE antibody has a heavy chain variableregion comprising SEQ ID NO: 113. In some embodiments, the heavy chainvariable region further comprises SEQ ID NO: 109 and/or SEQ ID NO: 111.In other embodiments, the heavy chain variable region further comprisesSEQ ID NO: 110 and/or SEQ ID NO: 112. In certain of the aboveembodiments, the antibody has a light chain variable region comprisingSEQ ID NO: 107. The light chain variable region can further comprise (a)SEQ ID NO: 105; and/or (b) SEQ ID NO: 106.

In another aspect, an anti-ApoE antibody has a heavy chain variableregion comprising SEQ ID NO: 114. In some embodiments, the heavy chainvariable region further comprises SEQ ID NO: 109 and/or SEQ ID NO: 111.In other embodiments, the heavy chain variable region further comprisesSEQ ID NO: 110 and/or SEQ ID NO: 112. In certain of the aboveembodiments, the antibody has a light chain variable region comprisingSEQ ID NO: 108. The light chain variable region can further comprise (a)SEQ ID NO: 105; and/or (b) SEQ ID NO: 106.

In another aspect, an anti-ApoE antibody is selected from Table E.

TABLE E Group III Antibodies Light Chain HVR Heavy Chain HVR Antibody L1L2 L3 H1 H2 H3 1 SEQ ID NO: 105 2 SEQ ID NO: 105 SEQ ID NO: 106 3 SEQ IDNO: 105 SEQ ID NO: 106 SEQ ID NO: 107 4 SEQ ID NO: 106 5 SEQ ID NO: 106SEQ ID NO: 107 6 SEQ ID NO: 107 7 SEQ ID NO: 105 SEQ ID NO: 107 8 SEQ IDNO: 109 9 SEQ ID NO: 109 SEQ ID NO: 111 10 SEQ ID NO: 109 SEQ ID NO: 111SEQ ID NO: 113 11 SEQ ID NO: 111 12 SEQ ID NO: 111 SEQ ID NO: 113 13 SEQID NO: 113 14 SEQ ID NO: 109 SEQ ID NO: 113 15 SEQ ID NO: 105 SEQ ID NO:109 16 SEQ ID NO: 105 SEQ ID NO: 109 SEQ ID NO: 111 17 SEQ ID NO: 105SEQ ID NO: 109 SEQ ID NO: 111 SEQ ID NO: 113 18 SEQ ID NO: 105 SEQ IDNO: 111 19 SEQ ID NO: 105 SEQ ID NO: 111 SEQ ID NO: 113 20 SEQ ID NO:105 SEQ ID NO: 113 21 SEQ ID NO: 105 SEQ ID NO: 109 SEQ ID NO: 113 22SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 109 23 SEQ ID NO: 105 SEQ IDNO: 106 SEQ ID NO: 109 SEQ ID NO: 111 24 SEQ ID NO: 105 SEQ ID NO: 106SEQ ID NO: 109 SEQ ID NO: 111 SEQ ID NO: 113 25 SEQ ID NO: 105 SEQ IDNO: 106 SEQ ID NO: 111 26 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 111SEQ ID NO: 113 27 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 113 28 SEQ IDNO: 105 SEQ ID NO: 106 SEQ ID NO: 109 SEQ ID NO: 113 29 SEQ ID NO: 105SEQ ID NO: 106 SEQ ID NO: 107 SEQ ID NO: 109 30 SEQ ID NO: 105 SEQ IDNO: 106 SEQ ID NO: 107 SEQ ID NO: 109 SEQ ID NO: 111 31 SEQ ID NO: 105SEQ ID NO: 106 SEQ ID NO: 107 SEQ ID NO: 109 SEQ ID NO: 111 SEQ ID NO:113 32 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 107 SEQ ID NO: 111 33SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 107 SEQ ID NO: 111 SEQ ID NO:113 34 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 107 SEQ ID NO: 109 SEQID NO: 113 35 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 107 SEQ ID NO:113 36 SEQ ID NO: 106 SEQ ID NO: 109 37 SEQ ID NO: 106 SEQ ID NO: 109SEQ ID NO: 111 38 SEQ ID NO: 106 SEQ ID NO: 109 SEQ ID NO: 111 SEQ IDNO: 113 39 SEQ ID NO: 106 SEQ ID NO: 111 40 SEQ ID NO: 106 SEQ ID NO:111 SEQ ID NO: 113 41 SEQ ID NO: 106 SEQ ID NO: 113 42 SEQ ID NO: 106SEQ ID NO: 109 SEQ ID NO: 113 43 SEQ ID NO: 106 SEQ ID NO: 107 SEQ IDNO: 109 44 SEQ ID NO: 106 SEQ ID NO: 107 SEQ ID NO: 109 SEQ ID NO: 11145 SEQ ID NO: 106 SEQ ID NO: 107 SEQ ID NO: 109 SEQ ID NO: 111 SEQ IDNO: 113 46 SEQ ID NO: 106 SEQ ID NO: 107 SEQ ID NO: 111 47 SEQ ID NO:106 SEQ ID NO: 107 SEQ ID NO: 111 SEQ ID NO: 113 48 SEQ ID NO: 106 SEQID NO: 107 SEQ ID NO: 113 49 SEQ ID NO: 106 SEQ ID NO: 107 SEQ ID NO:109 SEQ ID NO: 113 50 SEQ ID NO: 107 SEQ ID NO: 109 51 SEQ ID NO: 107SEQ ID NO: 109 SEQ ID NO: 111 52 SEQ ID NO: 107 SEQ ID NO: 109 SEQ IDNO: 111 SEQ ID NO: 113 53 SEQ ID NO: 107 SEQ ID NO: 111 54 SEQ ID NO:107 SEQ ID NO: 111 SEQ ID NO: 113 55 SEQ ID NO: 107 SEQ ID NO: 113 56SEQ ID NO: 107 SEQ ID NO: 109 SEQ ID NO: 113 57 SEQ ID NO: 105 SEQ IDNO: 107 SEQ ID NO: 109 58 SEQ ID NO: 105 SEQ ID NO: 107 SEQ ID NO: 109SEQ ID NO: 111 59 SEQ ID NO: 105 SEQ ID NO: 107 SEQ ID NO: 109 SEQ IDNO: 111 SEQ ID NO: 113 60 SEQ ID NO: 105 SEQ ID NO: 107 SEQ ID NO: 11161 SEQ ID NO: 105 SEQ ID NO: 107 SEQ ID NO: 111 SEQ ID NO: 113 62 SEQ IDNO: 105 SEQ ID NO: 107 SEQ ID NO: 113 63 SEQ ID NO: 105 SEQ ID NO: 107SEQ ID NO: 109 SEQ ID NO: 113 64 SEQ ID NO: 105 65 SEQ ID NO: 105 SEQ IDNO: 106 66 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 108 67 SEQ ID NO:106 68 SEQ ID NO: 106 SEQ ID NO: 108 69 SEQ ID NO: 108 70 SEQ ID NO: 105SEQ ID NO: 108 71 SEQ ID NO: 110 72 SEQ ID NO: 110 SEQ ID NO: 112 73 SEQID NO: 110 SEQ ID NO: 112 SEQ ID NO: 114 74 SEQ ID NO: 112 75 SEQ ID NO:112 SEQ ID NO: 114 76 SEQ ID NO: 114 77 SEQ ID NO: 110 SEQ ID NO: 114 78SEQ ID NO: 105 SEQ ID NO: 110 79 SEQ ID NO: 105 SEQ ID NO: 110 SEQ IDNO: 112 80 SEQ ID NO: 105 SEQ ID NO: 110 SEQ ID NO: 112 SEQ ID NO: 11481 SEQ ID NO: 105 SEQ ID NO: 112 82 SEQ ID NO: 105 SEQ ID NO: 112 SEQ IDNO: 114 83 SEQ ID NO: 105 SEQ ID NO: 114 84 SEQ ID NO: 105 SEQ ID NO:110 SEQ ID NO: 114 85 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 110 86SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 110 SEQ ID NO: 112 87 SEQ IDNO: 105 SEQ ID NO: 106 SEQ ID NO: 110 SEQ ID NO: 112 SEQ ID NO: 114 88SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 112 89 SEQ ID NO: 105 SEQ IDNO: 106 SEQ ID NO: 112 SEQ ID NO: 114 90 SEQ ID NO: 105 SEQ ID NO: 106SEQ ID NO: 114 91 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 110 SEQ IDNO: 114 92 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 108 SEQ ID NO: 11093 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 108 SEQ ID NO: 110 SEQ IDNO: 112 94 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 108 SEQ ID NO: 110SEQ ID NO: 112 SEQ ID NO: 114 95 SEQ ID NO: 105 SEQ ID NO: 106 SEQ IDNO: 108 SEQ ID NO: 112 96 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 108SEQ ID NO: 112 SEQ ID NO: 114 97 SEQ ID NO: 105 SEQ ID NO: 106 SEQ IDNO: 108 SEQ ID NO: 114 98 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 108SEQ ID NO: 110 SEQ ID NO: 114 99 SEQ ID NO: 106 SEQ ID NO: 110 100 SEQID NO: 106 SEQ ID NO: 110 SEQ ID NO: 112 101 SEQ ID NO: 106 SEQ ID NO:110 SEQ ID NO: 112 SEQ ID NO: 114 102 SEQ ID NO: 106 SEQ ID NO: 112 103SEQ ID NO: 106 SEQ ID NO: 112 SEQ ID NO: 114 104 SEQ ID NO: 106 SEQ IDNO: 114 105 SEQ ID NO: 106 SEQ ID NO: 110 SEQ ID NO: 114 106 SEQ ID NO:106 SEQ ID NO: 108 SEQ ID NO: 110 107 SEQ ID NO: 106 SEQ ID NO: 108 SEQID NO: 110 SEQ ID NO: 112 108 SEQ ID NO: 106 SEQ ID NO: 108 SEQ ID NO:110 SEQ ID NO: 112 SEQ ID NO: 114 109 SEQ ID NO: 106 SEQ ID NO: 108 SEQID NO: 112 110 SEQ ID NO: 106 SEQ ID NO: 108 SEQ ID NO: 112 SEQ ID NO:114 111 SEQ ID NO: 106 SEQ ID NO: 108 SEQ ID NO: 114 112 SEQ ID NO: 106SEQ ID NO: 108 SEQ ID NO: 110 SEQ ID NO: 114 113 SEQ ID NO: 108 SEQ IDNO: 110 114 SEQ ID NO: 108 SEQ ID NO: 110 SEQ ID NO: 112 115 SEQ ID NO:108 SEQ ID NO: 110 SEQ ID NO: 112 SEQ ID NO: 114 116 SEQ ID NO: 108 SEQID NO: 112 117 SEQ ID NO: 108 SEQ ID NO: 112 SEQ ID NO: 114 118 SEQ IDNO: 108 SEQ ID NO: 114 119 SEQ ID NO: 108 SEQ ID NO: 110 SEQ ID NO: 114120 SEQ ID NO: 105 SEQ ID NO: 108 SEQ ID NO: 110 121 SEQ ID NO: 105 SEQID NO: 108 SEQ ID NO: 110 SEQ ID NO: 112 122 SEQ ID NO: 105 SEQ ID NO:108 SEQ ID NO: 110 SEQ ID NO: 112 SEQ ID NO: 114 123 SEQ ID NO: 105 SEQID NO: 108 SEQ ID NO: 112 124 SEQ ID NO: 105 SEQ ID NO: 108 SEQ ID NO:112 SEQ ID NO: 114 125 SEQ ID NO: 105 SEQ ID NO: 108 SEQ ID NO: 114 126SEQ ID NO: 105 SEQ ID NO: 108 SEQ ID NO: 110 SEQ ID NO: 114

In an exemplary embodiment, an anti-ApoE antibody of this groupcomprises a VL that has one or more HVRs derived from SEQ ID NO: 115 ora VH that has one or more HVRs derived from SEQ ID NO: 116. The HVRderived from SEQ ID NO: 115 may be L1, L2, L3, or any combinationthereof. In certain embodiments, the VL may comprise an L1 of SEQ ID NO:105, an L2 of SEQ ID NO: 106, an L3 of SEQ ID NO: 117, or anycombination thereof (e.g. antibodies 1-7 in Table F). The HVR derivedfrom SEQ ID NO: 116 may be H1, H2, H3, or any combination thereof. Incertain embodiments, the VH may comprise an H1 of SEQ ID NO: 118, an H2of SEQ ID NO: 119, an H3 of SEQ ID NO: 120, or any combination thereof(e.g. antibodies 8-14 in Table F). The antibody comprising one or moreHVRs derived from SEQ ID NO: 116 may further comprise a light chainvariable region (VL) comprising one or more HVRs derived from SEQ ID NO:115. The HVR may be L1, L2, L3, or any combination thereof. In apreferred embodiment, the VL may comprise an L1 of SEQ ID NO: 105, an L2of SEQ ID NO: 106, an L3 of SEQ ID NO: 117, or any combination thereof(e.g. antibodies 15-63 in Table F). In various embodiments above, theantibody may be a humanized antibody, or the antibody may have a VL with90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% identity to SEQ ID NO:115 and/or a VH with 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100%identity to SEQ ID NO: 116. In each of the above embodiments, theanti-ApoE antibody may optionally comprise one or more constant regions,or a portion of a constant region, that is substantially human (i.e. atleast 85%, 90%, 95%, or 99% sequence identity with a known humanframework sequence). The present disclosure also encompasses thecorresponding nucleic acid sequences of SEQ ID NO: 115, 116, 105, 106,117, 118, 119, and 120, which can readily be determined by one of skillin the art, and may be incorporated into a vector or other large DNAmolecule, such as a chromosome, in order to express an antibody of thedisclosure.

In another exemplary embodiment, an anti-ApoE antibody of this groupcomprises a VL that has one or more HVRs derived from SEQ ID NO: 121 ora VH that has one or more HVRs derived from SEQ ID NO: 122. The HVRderived from SEQ ID NO: 121 may be L1, L2, L3, or any combinationthereof. In certain embodiments, the VL may comprise an L1 of SEQ ID NO:105, an L2 of SEQ ID NO: 106, an L3 of SEQ ID NO: 123, or anycombination thereof (e.g. antibodies 64-70 in Table F). The HVR derivedfrom SEQ ID NO: 122 may be H1, H2, H3, or any combination thereof. Incertain embodiments, the VH may comprise an H1 of SEQ ID NO: 124, an H2of SEQ ID NO: 125, an H3 of SEQ ID NO: 126, or any combination thereof(e.g. antibodies 71-77 in Table F). The antibody comprising one or moreHVRs derived from SEQ ID NO: 122 may further comprise a light chainvariable region (VL) comprising one or more HVRs derived from SEQ ID NO:121. The HVR may be L1, L2, L3, or any combination thereof. In apreferred embodiment, the VL may comprise an L1 of SEQ ID NO: 105, an L2of SEQ ID NO: 106, an L3 of SEQ ID NO: 123, or any combination thereof(e.g. antibodies 78-126 in Table F). In various embodiments above, theantibody may be a humanized antibody, or the antibody may have a VL with90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% identity to SEQ ID NO:121 and/or a VH with 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100%identity to SEQ ID NO: 122. In each of the above embodiments, theanti-ApoE antibody may optionally comprise one or more constant regions,or a portion of a constant region, that is substantially human (i.e. atleast 85%, 90%, 95%, or 99% sequence identity with a known humanframework sequence). The present disclosure also encompasses thecorresponding nucleic acid sequences of SEQ ID NO: 121, 122, 105, 106,123, 124, 125, and 126, which can readily be determined by one of skillin the art, and may be incorporated into a vector or other large DNAmolecule, such as a chromosome, in order to express an antibody of thedisclosure.

In some embodiments, each of the exemplary antibodies described abovemay also contain a variant Fc region, including but not limited to avariant Fc region that is modified to alter the natural interaction withthe microglia FcR.

In further embodiments, an isolated antibody of Group III recognizes anepitope listed in Tables 4-7.

TABLE F Exemplary Group III Antibodies Light Chain HVR Heavy Chain HVRAntibody L1 L2 L3 H1 H2 H3 1 SEQ ID NO: 105 2 SEQ ID NO: 105 SEQ ID NO:106 3 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 117 4 SEQ ID NO: 106 5SEQ ID NO: 106 SEQ ID NO: 117 6 SEQ ID NO: 117 7 SEQ ID NO: 105 SEQ IDNO: 117 8 SEQ ID NO: 118 9 SEQ ID NO: 118 SEQ ID NO: 119 10 SEQ ID NO:118 SEQ ID NO: 119 SEQ ID NO: 120 11 SEQ ID NO: 119 12 SEQ ID NO: 119SEQ ID NO: 120 13 SEQ ID NO: 120 14 SEQ ID NO: 118 SEQ ID NO: 120 15 SEQID NO: 105 SEQ ID NO: 118 16 SEQ ID NO: 105 SEQ ID NO: 118 SEQ ID NO:119 17 SEQ ID NO: 105 SEQ ID NO: 118 SEQ ID NO: 119 SEQ ID NO: 120 18SEQ ID NO: 105 SEQ ID NO: 119 19 SEQ ID NO: 105 SEQ ID NO: 119 SEQ IDNO: 120 20 SEQ ID NO: 105 SEQ ID NO: 120 21 SEQ ID NO: 105 SEQ ID NO:118 SEQ ID NO: 120 22 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 118 23SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 118 SEQ ID NO: 119 24 SEQ IDNO: 105 SEQ ID NO: 106 SEQ ID NO: 118 SEQ ID NO: 119 SEQ ID NO: 120 25SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 119 26 SEQ ID NO: 105 SEQ IDNO: 106 SEQ ID NO: 119 SEQ ID NO: 120 27 SEQ ID NO: 105 SEQ ID NO: 106SEQ ID NO: 120 28 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 118 SEQ IDNO: 120 29 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 117 SEQ ID NO: 11830 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 117 SEQ ID NO: 118 SEQ IDNO: 119 31 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 117 SEQ ID NO: 118SEQ ID NO: 119 SEQ ID NO: 120 32 SEQ ID NO: 105 SEQ ID NO: 106 SEQ IDNO: 117 SEQ ID NO: 119 33 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 117SEQ ID NO: 119 SEQ ID NO: 120 34 SEQ ID NO: 105 SEQ ID NO: 106 SEQ IDNO: 117 SEQ ID NO: 118 SEQ ID NO: 120 35 SEQ ID NO: 105 SEQ ID NO: 106SEQ ID NO: 117 SEQ ID NO: 120 36 SEQ ID NO: 106 SEQ ID NO: 118 37 SEQ IDNO: 106 SEQ ID NO: 118 SEQ ID NO: 119 38 SEQ ID NO: 106 SEQ ID NO: 118SEQ ID NO: 119 SEQ ID NO: 120 39 SEQ ID NO: 106 SEQ ID NO: 119 40 SEQ IDNO: 106 SEQ ID NO: 119 SEQ ID NO: 120 41 SEQ ID NO: 106 SEQ ID NO: 12042 SEQ ID NO: 106 SEQ ID NO: 118 SEQ ID NO: 120 43 SEQ ID NO: 106 SEQ IDNO: 117 SEQ ID NO: 118 44 SEQ ID NO: 106 SEQ ID NO: 117 SEQ ID NO: 118SEQ ID NO: 119 45 SEQ ID NO: 106 SEQ ID NO: 117 SEQ ID NO: 118 SEQ IDNO: 119 SEQ ID NO: 120 46 SEQ ID NO: 106 SEQ ID NO: 117 SEQ ID NO: 11947 SEQ ID NO: 106 SEQ ID NO: 117 SEQ ID NO: 119 SEQ ID NO: 120 48 SEQ IDNO: 106 SEQ ID NO: 117 SEQ ID NO: 120 49 SEQ ID NO: 106 SEQ ID NO: 117SEQ ID NO: 118 SEQ ID NO: 120 50 SEQ ID NO: 117 SEQ ID NO: 118 51 SEQ IDNO: 117 SEQ ID NO: 118 SEQ ID NO: 119 52 SEQ ID NO: 117 SEQ ID NO: 118SEQ ID NO: 119 SEQ ID NO: 120 53 SEQ ID NO: 117 SEQ ID NO: 119 54 SEQ IDNO: 117 SEQ ID NO: 119 SEQ ID NO: 120 55 SEQ ID NO: 117 SEQ ID NO: 12056 SEQ ID NO: 117 SEQ ID NO: 118 SEQ ID NO: 120 57 SEQ ID NO: 105 SEQ IDNO: 117 SEQ ID NO: 118 58 SEQ ID NO: 105 SEQ ID NO: 117 SEQ ID NO: 118SEQ ID NO: 119 59 SEQ ID NO: 105 SEQ ID NO: 117 SEQ ID NO: 118 SEQ IDNO: 119 SEQ ID NO: 120 60 SEQ ID NO: 105 SEQ ID NO: 117 SEQ ID NO: 11961 SEQ ID NO: 105 SEQ ID NO: 117 SEQ ID NO: 119 SEQ ID NO: 120 62 SEQ IDNO: 105 SEQ ID NO: 117 SEQ ID NO: 120 63 SEQ ID NO: 105 SEQ ID NO: 117SEQ ID NO: 118 SEQ ID NO: 120 64 SEQ ID NO: 105 65 SEQ ID NO: 105 SEQ IDNO: 106 66 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 123 67 SEQ ID NO:106 68 SEQ ID NO: 106 SEQ ID NO: 123 69 SEQ ID NO: 123 70 SEQ ID NO: 105SEQ ID NO: 123 71 SEQ ID NO: 124 72 SEQ ID NO: 124 SEQ ID NO: 125 73 SEQID NO: 124 SEQ ID NO: 125 SEQ ID NO: 126 74 SEQ ID NO: 125 75 SEQ ID NO:125 SEQ ID NO: 126 76 SEQ ID NO: 126 77 SEQ ID NO: 124 SEQ ID NO: 126 78SEQ ID NO: 105 SEQ ID NO: 124 79 SEQ ID NO: 105 SEQ ID NO: 124 SEQ IDNO: 125 80 SEQ ID NO: 105 SEQ ID NO: 124 SEQ ID NO: 125 SEQ ID NO: 12681 SEQ ID NO: 105 SEQ ID NO: 125 82 SEQ ID NO: 105 SEQ ID NO: 125 SEQ IDNO: 126 83 SEQ ID NO: 105 SEQ ID NO: 126 84 SEQ ID NO: 105 SEQ ID NO:124 SEQ ID NO: 126 85 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 124 86SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 124 SEQ ID NO: 125 87 SEQ IDNO: 105 SEQ ID NO: 106 SEQ ID NO: 124 SEQ ID NO: 125 SEQ ID NO: 126 88SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 125 89 SEQ ID NO: 105 SEQ IDNO: 106 SEQ ID NO: 125 SEQ ID NO: 126 90 SEQ ID NO: 105 SEQ ID NO: 106SEQ ID NO: 126 91 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 124 SEQ IDNO: 126 92 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 123 SEQ ID NO: 12493 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 123 SEQ ID NO: 124 SEQ IDNO: 125 94 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 123 SEQ ID NO: 124SEQ ID NO: 125 SEQ ID NO: 126 95 SEQ ID NO: 105 SEQ ID NO: 106 SEQ IDNO: 123 SEQ ID NO: 125 96 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 123SEQ ID NO: 125 SEQ ID NO: 126 97 SEQ ID NO: 105 SEQ ID NO: 106 SEQ IDNO: 123 SEQ ID NO: 126 98 SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO: 123SEQ ID NO: 124 SEQ ID NO: 126 99 SEQ ID NO: 106 SEQ ID NO: 124 100 SEQID NO: 106 SEQ ID NO: 124 SEQ ID NO: 125 101 SEQ ID NO: 106 SEQ ID NO:124 SEQ ID NO: 125 SEQ ID NO: 126 102 SEQ ID NO: 106 SEQ ID NO: 125 103SEQ ID NO: 106 SEQ ID NO: 125 SEQ ID NO: 126 104 SEQ ID NO: 106 SEQ IDNO: 126 105 SEQ ID NO: 106 SEQ ID NO: 124 SEQ ID NO: 126 106 SEQ ID NO:106 SEQ ID NO: 123 SEQ ID NO: 124 107 SEQ ID NO: 106 SEQ ID NO: 123 SEQID NO: 124 SEQ ID NO: 125 108 SEQ ID NO: 106 SEQ ID NO: 123 SEQ ID NO:124 SEQ ID NO: 125 SEQ ID NO: 126 109 SEQ ID NO: 106 SEQ ID NO: 123 SEQID NO: 125 110 SEQ ID NO: 106 SEQ ID NO: 123 SEQ ID NO: 125 SEQ ID NO:126 111 SEQ ID NO: 106 SEQ ID NO: 123 SEQ ID NO: 126 112 SEQ ID NO: 106SEQ ID NO: 123 SEQ ID NO: 124 SEQ ID NO: 126 113 SEQ ID NO: 123 SEQ IDNO: 124 114 SEQ ID NO: 123 SEQ ID NO: 124 SEQ ID NO: 125 115 SEQ ID NO:123 SEQ ID NO: 124 SEQ ID NO: 125 SEQ ID NO: 126 116 SEQ ID NO: 123 SEQID NO: 125 117 SEQ ID NO: 123 SEQ ID NO: 125 SEQ ID NO: 126 118 SEQ IDNO: 123 SEQ ID NO: 126 119 SEQ ID NO: 123 SEQ ID NO: 124 SEQ ID NO: 126120 SEQ ID NO: 105 SEQ ID NO: 123 SEQ ID NO: 124 121 SEQ ID NO: 105 SEQID NO: 123 SEQ ID NO: 124 SEQ ID NO: 125 122 SEQ ID NO: 105 SEQ ID NO:123 SEQ ID NO: 124 SEQ ID NO: 125 SEQ ID NO: 126 123 SEQ ID NO: 105 SEQID NO: 123 SEQ ID NO: 125 124 SEQ ID NO: 105 SEQ ID NO: 123 SEQ ID NO:125 SEQ ID NO: 126 125 SEQ ID NO: 105 SEQ ID NO: 123 SEQ ID NO: 126 126SEQ ID NO: 105 SEQ ID NO: 123 SEQ ID NO: 124 SEQ ID NO: 126(d) Anti-ApoE Antibodies that Preferentially Bind to ApoE4 Over ApoE3 orApoE2

In exemplary embodiment, an anti-ApoE antibody of this group comprises aVL that has one or more HVRs derived from SEQ ID NO: 1 or a VH that hasone or more HVRs derived from SEQ ID NO: 2. The HVR derived from SEQ IDNO: 1 may be L1, L2, L3, or any combination thereof. In certainembodiments, the VL may comprise an L1 of SEQ ID NO: 23, an L2 of SEQ IDNO: 24, an L3 of SEQ ID NO: 96, or any combination thereof (e.g.antibodies 1-7 in Table G). The HVR derived from SEQ ID NO: 2 may be H1,H2, H3, or any combination thereof. In certain embodiments, the VH maycomprise an H1 of SEQ ID NO: 26, an H2 of SEQ ID NO: 27, an H3 of SEQ IDNO: 28, or any combination thereof (e.g. antibodies 8-14 in Table G).The antibody comprising one or more HVRs derived from SEQ ID NO: 2 mayfurther comprise a light chain variable region (VL) comprising one ormore HVRs derived from SEQ ID NO: 1. The HVR may be L1, L2, L3, or anycombination thereof. In a preferred embodiment, the VL may comprise anL1 of SEQ ID NO: 23, an L2 of SEQ ID NO: 24, an L3 of SEQ ID NO: 96, orany combination thereof (e.g. antibodies 15-63 in Table G). In variousembodiments above, the antibody may be a humanized antibody, or theantibody may have a VL with 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or100% identity to SEQ ID NO: 1 and/or a VH with 90, 91, 92, 93, 94, 95,96, 97, 98, 99 or 100% identity to SEQ ID NO: 2. In each of the aboveembodiments, the anti-ApoE antibody may optionally comprise one or moreconstant regions, or a portion of a constant region, that issubstantially human (i.e. at least 90%, 95%, or 99% sequence identitywith a known human framework sequence). The present disclosure alsoencompasses the corresponding nucleic acid sequences of SEQ ID NO: 1, 2,23, 24, 96, 26, 27, and 28, which can readily be determined by one ofskill in the art, and may be incorporated into a vector or other largeDNA molecule, such as a chromosome, in order to express an antibody ofthe disclosure.

In some embodiments, each of the exemplary antibodies described abovemay also contain a variant Fc region, including but not limited to avariant Fc region that is modified to alter the natural interaction withthe microglia FcR.

In further embodiments, an isolated antibody of Group IV recognizes anepitope listed in Tables 4-7, for example, as described for theexemplary antibody HJ151.

TABLE G Exemplary Antibodies Light Chain HVR Heavy Chain HVR Antibody L1L2 L3 H1 H2 H3 1 SEQ ID NO: 23 2 SEQ ID NO: 23 SEQ ID NO: 24 3 SEQ IDNO: 23 SEQ ID NO: 24 SEQ ID NO: 96 4 SEQ ID NO: 24 5 SEQ ID NO: 24 SEQID NO: 96 6 SEQ ID NO: 96 7 SEQ ID NO: 23 SEQ ID NO: 96 8 SEQ ID NO: 269 SEQ ID NO: 26 SEQ ID NO: 27 10 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO:28 11 SEQ ID NO: 27 12 SEQ ID NO: 27 SEQ ID NO: 28 13 SEQ ID NO: 28 14SEQ ID NO: 26 SEQ ID NO: 28 15 SEQ ID NO: 23 SEQ ID NO: 26 16 SEQ ID NO:23 SEQ ID NO: 26 SEQ ID NO: 27 17 SEQ ID NO: 23 SEQ ID NO: 26 SEQ ID NO:27 SEQ ID NO: 28 18 SEQ ID NO: 23 SEQ ID NO: 27 19 SEQ ID NO: 23 SEQ IDNO: 27 SEQ ID NO: 28 20 SEQ ID NO: 23 SEQ ID NO: 28 21 SEQ ID NO: 23 SEQID NO: 26 SEQ ID NO: 28 22 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 26 23SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 26 SEQ ID NO: 27 24 SEQ ID NO: 23SEQ ID NO: 24 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 25 SEQ ID NO: 23SEQ ID NO: 24 SEQ ID NO: 27 26 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 27SEQ ID NO: 28 27 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 28 28 SEQ ID NO:23 SEQ ID NO: 24 SEQ ID NO: 26 SEQ ID NO: 28 29 SEQ ID NO: 23 SEQ ID NO:24 SEQ ID NO: 96 SEQ ID NO: 26 30 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO:96 SEQ ID NO: 26 SEQ ID NO: 27 31 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO:96 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 32 SEQ ID NO: 23 SEQ ID NO:24 SEQ ID NO: 96 SEQ ID NO: 27 33 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO:96 SEQ ID NO: 27 SEQ ID NO: 28 34 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO:96 SEQ ID NO: 26 SEQ ID NO: 28 35 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO:96 SEQ ID NO: 28 36 SEQ ID NO: 24 SEQ ID NO: 26 37 SEQ ID NO: 24 SEQ IDNO: 26 SEQ ID NO: 27 38 SEQ ID NO: 24 SEQ ID NO: 26 SEQ ID NO: 27 SEQ IDNO: 28 39 SEQ ID NO: 24 SEQ ID NO: 27 40 SEQ ID NO: 24 SEQ ID NO: 27 SEQID NO: 28 41 SEQ ID NO: 24 SEQ ID NO: 28 42 SEQ ID NO: 24 SEQ ID NO: 26SEQ ID NO: 28 43 SEQ ID NO: 24 SEQ ID NO: 96 SEQ ID NO: 26 44 SEQ ID NO:24 SEQ ID NO: 96 SEQ ID NO: 26 SEQ ID NO: 27 45 SEQ ID NO: 24 SEQ ID NO:96 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 46 SEQ ID NO: 24 SEQ ID NO:96 SEQ ID NO: 27 47 SEQ ID NO: 24 SEQ ID NO: 96 SEQ ID NO: 27 SEQ ID NO:28 48 SEQ ID NO: 24 SEQ ID NO: 96 SEQ ID NO: 28 49 SEQ ID NO: 24 SEQ IDNO: 96 SEQ ID NO: 26 SEQ ID NO: 28 50 SEQ ID NO: 96 SEQ ID NO: 26 51 SEQID NO: 96 SEQ ID NO: 26 SEQ ID NO: 27 52 SEQ ID NO: 96 SEQ ID NO: 26 SEQID NO: 27 SEQ ID NO: 28 53 SEQ ID NO: 96 SEQ ID NO: 27 54 SEQ ID NO: 96SEQ ID NO: 27 SEQ ID NO: 28 55 SEQ ID NO: 96 SEQ ID NO: 28 56 SEQ ID NO:96 SEQ ID NO: 26 SEQ ID NO: 28 57 SEQ ID NO: 23 SEQ ID NO: 96 SEQ ID NO:26 58 SEQ ID NO: 23 SEQ ID NO: 96 SEQ ID NO: 26 SEQ ID NO: 27 59 SEQ IDNO: 23 SEQ ID NO: 96 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 60 SEQ IDNO: 23 SEQ ID NO: 96 SEQ ID NO: 27 61 SEQ ID NO: 23 SEQ ID NO: 96 SEQ IDNO: 27 SEQ ID NO: 28 62 SEQ ID NO: 23 SEQ ID NO: 96 SEQ ID NO: 28 63 SEQID NO: 23 SEQ ID NO: 96 SEQ ID NO: 26 SEQ ID NO: 28(e) Additional Anti-ApoE Antibodies that do not Preferentially Bind toApoE2, ApoE3 or ApoE4

In an exemplary embodiment, an anti-ApoE antibody of this groupcomprises a VL that has one or more HVRs derived from SEQ ID NO: 21 or aVH that has one or more HVRs derived from SEQ ID NO: 22. The HVR derivedfrom SEQ ID NO: 21 may be L1, L2, L3, or any combination thereof. Incertain embodiments, the VL may comprise an L1 of SEQ ID NO: 72, an L2of SEQ ID NO: 73, an L3 of SEQ ID NO: 74, or any combination thereof(e.g. antibodies 1-7 in Table H). The HVR derived from SEQ ID NO: 22 maybe H1, H2, H3, or any combination thereof. In certain embodiments, theVH may comprise an H1 of SEQ ID NO: 75, an H2 of SEQ ID NO: 76, an H3 ofSEQ ID NO: 77, or any combination thereof (e.g. antibodies 8-14 in TableH). The antibody comprising one or more HVRs derived from SEQ ID NO: 22may further comprise a light chain variable region (VL) comprising oneor more HVRs derived from SEQ ID NO: 21. The HVR may be L1, L2, L3, orany combination thereof. In a preferred embodiment, the VL may comprisean L1 of SEQ ID NO: 72, an L2 of SEQ ID NO: 73, an L3 of SEQ ID NO: 74,or any combination thereof (e.g. antibodies 15-63 in Table H). Invarious embodiments above, the antibody may be a humanized antibody, orthe antibody may have a VL with 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,or 100% identity to SEQ ID NO: 21 and/or a VH with 90, 91, 92, 93, 94,95, 96, 97, 98, 99 or 100% identity to SEQ ID NO: 22. In each of theabove embodiments, the anti-ApoE antibody may optionally comprise one ormore constant regions, or a portion of a constant region, that issubstantially human (i.e. at least 90%, 95%, or 99% sequence identitywith a known human framework sequence). The present disclosure alsoencompasses the corresponding nucleic acid sequences of SEQ ID NO: 21,22, 72, 73, 74, 75, 76, and 77, which can readily be determined by oneof skill in the art, and may be incorporated into a vector or otherlarge DNA molecule, such as a chromosome, in order to express anantibody of the disclosure.

In an exemplary embodiment, an anti-ApoE antibody of this groupcomprises a VL that has one or more HVRs derived from SEQ ID NO: 97 or aVH that has one or more HVRs derived from SEQ ID NO: 98. The HVR derivedfrom SEQ ID NO: 97 may be L1, L2, L3, or any combination thereof. Incertain embodiments, the VL may comprise an L1 of SEQ ID NO: 99, an L2of SEQ ID NO: 100, an L3 of SEQ ID NO: 101, or any combination thereof(e.g. antibodies 64-70 in Table H). The HVR derived from SEQ ID NO: 98may be H1, H2, H3, or any combination thereof. In certain embodiments,the VH may comprise an H1 of SEQ ID NO: 102, an H2 of SEQ ID NO: 103, anH3 of SEQ ID NO: 104, or any combination thereof (e.g. antibodies 71-77in Table H). The antibody comprising one or more HVRs derived from SEQID NO: 98 may further comprise a light chain variable region (VL)comprising one or more HVRs derived from SEQ ID NO: 97. The HVR may beL1, L2, L3, or any combination thereof. In a preferred embodiment, theVL may comprise an L1 of SEQ ID NO: 99, an L2 of SEQ ID NO: 100, an L3of SEQ ID NO: 101, or any combination thereof (e.g. antibodies 78-126 inTable H). In various embodiments above, the antibody may be a humanizedantibody, or the antibody may have a VL with 90, 91, 92, 93, 94, 95, 96,97, 98, 99, or 100% identity to SEQ ID NO: 97 and/or a VH with 90, 91,92, 93, 94, 95, 96, 97, 98, 99 or 100% identity to SEQ ID NO: 98. Ineach of the above embodiments, the anti-ApoE antibody may optionallycomprise one or more constant regions, or a portion of a constantregion, that is substantially human (i.e. at least 90%, 95%, or 99%sequence identity with a known human framework sequence). The presentdisclosure also encompasses the corresponding nucleic acid sequences ofSEQ ID NO: 97, 98, 99, 100, 101, 102, 103, and 104, which can readily bedetermined by one of skill in the art, and may be incorporated into avector or other large DNA molecule, such as a chromosome, in order toexpress an antibody of the disclosure.

In some embodiments, each of the exemplary antibodies described abovemay also contain a variant Fc region, including but not limited to avariant Fc region that is modified to alter the natural interaction withthe microglia FcR.

In further embodiments, an isolated antibody of Group V recognizes anepitope listed in Tables 4-7, for example, as described for theexemplary antibody HJ1526.

TABLE H Exemplary Antibodies Light Chain HVR Heavy Chain HVR Antibody L1L2 L3 H1 H2 H3 1 SEQ ID NO: 72 2 SEQ ID NO: 72 SEQ ID NO: 73 3 SEQ IDNO: 72 SEQ ID NO: 73 SEQ ID NO: 74 4 SEQ ID NO: 73 5 SEQ ID NO: 73 SEQID NO: 74 6 SEQ ID NO: 74 7 SEQ ID NO: 72 SEQ ID NO: 74 8 SEQ ID NO: 759 SEQ ID NO: 75 SEQ ID NO: 76 10 SEQ ID NO: 75 SEQ ID NO: 76 SEQ ID NO:77 11 SEQ ID NO: 76 12 SEQ ID NO: 76 SEQ ID NO: 77 13 SEQ ID NO: 77 14SEQ ID NO: 75 SEQ ID NO: 77 15 SEQ ID NO: 72 SEQ ID NO: 75 16 SEQ ID NO:72 SEQ ID NO: 75 SEQ ID NO: 76 17 SEQ ID NO: 72 SEQ ID NO: 75 SEQ ID NO:76 SEQ ID NO: 77 18 SEQ ID NO: 72 SEQ ID NO: 76 19 SEQ ID NO: 72 SEQ IDNO: 76 SEQ ID NO: 77 20 SEQ ID NO: 72 SEQ ID NO: 77 21 SEQ ID NO: 72 SEQID NO: 75 SEQ ID NO: 77 22 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 75 23SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 75 SEQ ID NO: 76 24 SEQ ID NO: 72SEQ ID NO: 73 SEQ ID NO: 75 SEQ ID NO: 76 SEQ ID NO: 77 25 SEQ ID NO: 72SEQ ID NO: 73 SEQ ID NO: 76 26 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 76SEQ ID NO: 77 27 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 77 28 SEQ ID NO:72 SEQ ID NO: 73 SEQ ID NO: 75 SEQ ID NO: 77 29 SEQ ID NO: 72 SEQ ID NO:73 SEQ ID NO: 74 SEQ ID NO: 75 30 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO:74 SEQ ID NO: 75 SEQ ID NO: 76 31 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO:74 SEQ ID NO: 75 SEQ ID NO: 76 SEQ ID NO: 77 32 SEQ ID NO: 72 SEQ ID NO:73 SEQ ID NO: 74 SEQ ID NO: 76 33 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO:74 SEQ ID NO: 76 SEQ ID NO: 77 34 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO:74 SEQ ID NO: 75 SEQ ID NO: 77 35 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO:74 SEQ ID NO: 77 36 SEQ ID NO: 73 SEQ ID NO: 75 37 SEQ ID NO: 73 SEQ IDNO: 75 SEQ ID NO: 76 38 SEQ ID NO: 73 SEQ ID NO: 75 SEQ ID NO: 76 SEQ IDNO: 77 39 SEQ ID NO: 73 SEQ ID NO: 76 40 SEQ ID NO: 73 SEQ ID NO: 76 SEQID NO: 77 41 SEQ ID NO: 73 SEQ ID NO: 77 42 SEQ ID NO: 73 SEQ ID NO: 75SEQ ID NO: 77 43 SEQ ID NO: 73 SEQ ID NO: 74 SEQ ID NO: 75 44 SEQ ID NO:73 SEQ ID NO: 74 SEQ ID NO: 75 SEQ ID NO: 76 45 SEQ ID NO: 73 SEQ ID NO:74 SEQ ID NO: 75 SEQ ID NO: 76 SEQ ID NO: 77 46 SEQ ID NO: 73 SEQ ID NO:74 SEQ ID NO: 76 47 SEQ ID NO: 73 SEQ ID NO: 74 SEQ ID NO: 76 SEQ ID NO:77 48 SEQ ID NO: 73 SEQ ID NO: 74 SEQ ID NO: 77 49 SEQ ID NO: 73 SEQ IDNO: 74 SEQ ID NO: 75 SEQ ID NO: 77 50 SEQ ID NO: 74 SEQ ID NO: 75 51 SEQID NO: 74 SEQ ID NO: 75 SEQ ID NO: 76 52 SEQ ID NO: 74 SEQ ID NO: 75 SEQID NO: 76 SEQ ID NO: 77 53 SEQ ID NO: 74 SEQ ID NO: 76 54 SEQ ID NO: 74SEQ ID NO: 76 SEQ ID NO: 77 55 SEQ ID NO: 74 SEQ ID NO: 77 56 SEQ ID NO:74 SEQ ID NO: 75 SEQ ID NO: 77 57 SEQ ID NO: 72 SEQ ID NO: 74 SEQ ID NO:75 58 SEQ ID NO: 72 SEQ ID NO: 74 SEQ ID NO: 75 SEQ ID NO: 76 59 SEQ IDNO: 72 SEQ ID NO: 74 SEQ ID NO: 75 SEQ ID NO: 76 SEQ ID NO: 77 60 SEQ IDNO: 72 SEQ ID NO: 74 SEQ ID NO: 76 61 SEQ ID NO: 72 SEQ ID NO: 74 SEQ IDNO: 76 SEQ ID NO: 77 62 SEQ ID NO: 72 SEQ ID NO: 74 SEQ ID NO: 77 63 SEQID NO: 72 SEQ ID NO: 74 SEQ ID NO: 75 SEQ ID NO: 77 64 SEQ ID NO: 99 65SEQ ID NO: 99 SEQ ID NO: 100 66 SEQ ID NO: 99 SEQ ID NO: 100 SEQ ID NO:101 67 SEQ ID NO: 100 68 SEQ ID NO: 100 SEQ ID NO: 101 69 SEQ ID NO: 10170 SEQ ID NO: 99 SEQ ID NO: 101 71 SEQ ID NO: 102 72 SEQ ID NO: 102 SEQID NO: 103 73 SEQ ID NO: 102 SEQ ID NO: 103 SEQ ID NO: 104 74 SEQ ID NO:103 75 SEQ ID NO: 103 SEQ ID NO: 104 76 SEQ ID NO: 104 77 SEQ ID NO: 102SEQ ID NO: 104 78 SEQ ID NO: 99 SEQ ID NO: 102 79 SEQ ID NO: 99 SEQ IDNO: 102 SEQ ID NO: 103 80 SEQ ID NO: 99 SEQ ID NO: 102 SEQ ID NO: 103SEQ ID NO: 104 81 SEQ ID NO: 99 SEQ ID NO: 103 82 SEQ ID NO: 99 SEQ IDNO: 103 SEQ ID NO: 104 83 SEQ ID NO: 99 SEQ ID NO: 104 84 SEQ ID NO: 99SEQ ID NO: 102 SEQ ID NO: 104 85 SEQ ID NO: 99 SEQ ID NO: 100 SEQ ID NO:102 86 SEQ ID NO: 99 SEQ ID NO: 100 SEQ ID NO: 102 SEQ ID NO: 103 87 SEQID NO: 99 SEQ ID NO: 100 SEQ ID NO: 102 SEQ ID NO: 103 SEQ ID NO: 104 88SEQ ID NO: 99 SEQ ID NO: 100 SEQ ID NO: 103 89 SEQ ID NO: 99 SEQ ID NO:100 SEQ ID NO: 103 SEQ ID NO: 104 90 SEQ ID NO: 99 SEQ ID NO: 100 SEQ IDNO: 104 91 SEQ ID NO: 99 SEQ ID NO: 100 SEQ ID NO: 102 SEQ ID NO: 104 92SEQ ID NO: 99 SEQ ID NO: 100 SEQ ID NO: 101 SEQ ID NO: 102 93 SEQ ID NO:99 SEQ ID NO: 100 SEQ ID NO: 101 SEQ ID NO: 102 SEQ ID NO: 103 94 SEQ IDNO: 99 SEQ ID NO: 100 SEQ ID NO: 101 SEQ ID NO: 102 SEQ ID NO: 103 SEQID NO: 104 95 SEQ ID NO: 99 SEQ ID NO: 100 SEQ ID NO: 101 SEQ ID NO: 10396 SEQ ID NO: 99 SEQ ID NO: 100 SEQ ID NO: 101 SEQ ID NO: 103 SEQ ID NO:104 97 SEQ ID NO: 99 SEQ ID NO: 100 SEQ ID NO: 101 SEQ ID NO: 102 SEQ IDNO: 104 98 SEQ ID NO: 99 SEQ ID NO: 100 SEQ ID NO: 101 SEQ ID NO: 104 99SEQ ID NO: 100 SEQ ID NO: 102 100 SEQ ID NO: 100 SEQ ID NO: 102 SEQ IDNO: 103 101 SEQ ID NO: 100 SEQ ID NO: 102 SEQ ID NO: 103 SEQ ID NO: 104102 SEQ ID NO: 100 SEQ ID NO: 103 103 SEQ ID NO: 100 SEQ ID NO: 103 SEQID NO: 104 104 SEQ ID NO: 100 SEQ ID NO: 104 105 SEQ ID NO: 100 SEQ IDNO: 102 SEQ ID NO: 104 106 SEQ ID NO: 100 SEQ ID NO: 101 SEQ ID NO: 102107 SEQ ID NO: 100 SEQ ID NO: 101 SEQ ID NO: 102 SEQ ID NO: 103 108 SEQID NO: 100 SEQ ID NO: 101 SEQ ID NO: 102 SEQ ID NO: 103 SEQ ID NO: 104109 SEQ ID NO: 100 SEQ ID NO: 101 SEQ ID NO: 103 110 SEQ ID NO: 100 SEQID NO: 101 SEQ ID NO: 103 SEQ ID NO: 104 111 SEQ ID NO: 100 SEQ ID NO:101 SEQ ID NO: 104 112 SEQ ID NO: 100 SEQ ID NO: 101 SEQ ID NO: 102 SEQID NO: 104 113 SEQ ID NO: 101 SEQ ID NO: 102 114 SEQ ID NO: 101 SEQ IDNO: 102 SEQ ID NO: 103 115 SEQ ID NO: 101 SEQ ID NO: 102 SEQ ID NO: 103SEQ ID NO: 104 116 SEQ ID NO: 101 SEQ ID NO: 103 117 SEQ ID NO: 101 SEQID NO: 103 SEQ ID NO: 104 118 SEQ ID NO: 101 SEQ ID NO: 104 119 SEQ IDNO: 101 SEQ ID NO: 102 SEQ ID NO: 104 120 SEQ ID NO: 99 SEQ ID NO: 101SEQ ID NO: 102 121 SEQ ID NO: 99 SEQ ID NO: 101 SEQ ID NO: 102 SEQ IDNO: 103 122 SEQ ID NO: 99 SEQ ID NO: 101 SEQ ID NO: 102 SEQ ID NO: 103SEQ ID NO: 104 123 SEQ ID NO: 99 SEQ ID NO: 101 SEQ ID NO: 103 124 SEQID NO: 99 SEQ ID NO: 101 SEQ ID NO: 103 SEQ ID NO: 104 125 SEQ ID NO: 99SEQ ID NO: 101 SEQ ID NO: 104 126 SEQ ID NO: 99 SEQ ID NO: 101 SEQ IDNO: 102 SEQ ID NO: 104

(f) Group V

In another aspect, an anti-ApoE antibody competitively inhibits bindingof a reference antibody to its epitope. An antibody is said tocompetitively inhibit binding of a reference antibody to a given epitopeif the antibody preferentially binds to that epitope to the extent thatit blocks binding of the reference antibody to the epitope by at least50%, at least 60%, at least 70%, at least 80%, or at least 90%.Competitive inhibition can be determined by any method known in the art,for example, competition ELISA assays. In some embodiments, an anti-ApoEantibody competitively inhibits HJ152 and/or HJ1514 binding to humanApoE. In other embodiments, an anti-ApoE antibody competitively inhibitsHJ155, HJ156, HJ159, HJ1513 and/or HJ1518 binding to human ApoE. Instill other embodiments, an anti-ApoE antibody competitively inhibitsHJ151 binding to human ApoE. In yet other embodiments, an anti-ApoEantibody competitively inhibits HJ1526 binding to human ApoE. In analternative embodiment, an anti-ApoE antibody competitively inhibitsHJ158 binding to human ApoE. In a different embodiment, an anti-ApoEantibody competitively inhibits HJ1531 and/or HJ1536 binding to humanApoE.

III. Treatment Methods

The present disclosure provides a method of treating Aβ amyloidosis, themethod comprising administering a therapeutically effective amount of ananti-ApoE antibody to a subject in need thereof. The present disclosurealso provides a method of treating a subject diagnosed with a diseasecharacterized by brain Aβ plaques, the method comprising administering atherapeutically effective amount of an anti-ApoE antibody to thesubject. The present disclosure also provides a method of treating asubject diagnosed with a disease characterized by vascular Aβ plaques inthe brain, the method comprising administering a therapeuticallyeffective amount of an anti-ApoE antibody to the subject. The presentdisclosure also provides a method of preventing the progression of adisease characterized by Aβ plaques in the brain, the method comprisingadministering a therapeutically effective amount of an anti-ApoEantibody to a subject in need thereof. The present disclosure alsoprovides a method of treating a subject diagnosed with Alzheimer'sdisease, the method comprising administering a therapeutically effectiveamount of an anti-ApoE antibody to the subject. The present disclosurealso provides a method of treating a subject diagnosed with CAA, themethod comprising administering a therapeutically effective amount of ananti-ApoE antibody to the subject. Suitable anti-ApoE antibodies aredescribed in Section II. In embodiments where the subject is a human,the anti-ApoE antibody is adapted for administration to a living humansubject (e.g. humanized).

In one embodiment, the disclosure provides a method of preventing theprogression, or slowing the rate of progression, of a diseasecharacterized by Aβ plaques in the brain. The method comprisesadministering a therapeutically effective amount of an anti-ApoEantibody to a subject in need thereof. Suitable anti-ApoE antibodiesinclude those disclosed herein. Progression of a disease characterizedby Aβ plaques in the brain can be evaluated by methods known in the artand described herein, including a worsening of a clinical sign of Aβamyloidosis, an Aβ plaque associated symptom, or a CAA associatedsymptom. In exemplary embodiments, the clinical sign is amyloid plaqueload.

In another embodiment, the disclosure provides a method for improving aclinical sign of Aβ amyloidosis. The method comprises administering atherapeutically effective amount of an anti-ApoE antibody to a subjectin need thereof. Suitable anti-ApoE antibodies include those disclosedherein. Non-limiting examples of improved clinical signs of Aβamyloidosis may include a decrease in amyloid plaque load, stabilizationof amyloid plaque load (i.e. no further increase), an increase in CSFAβ42 concentration, an increase in CSF Aβ42/Aβ40 ratio, a decreasedAβ42/Aβ40 peak time ratio as measured by stable isotope labelingkinetics (e.g. such that is closer to 1), a decreased Aβ42/Aβ40 FTRratio as measured by stable isotope labeling kinetics (e.g. such that iscloser to 1), and a change in the ratio of the relative labeling Aβ42 tothe relative labeling of Aβ40 (or another Aβ peptide) after stableisotope labeling such that the ratio is closer to 1. In each of theabove embodiments, the improvement (i.e. the change) in the clinicalsign is at least statistically significant. In certain embodiments, thechange may be at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, or 20% compared to untreated or negative controltreated subjects. In some embodiments, the change may be at least 20,25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95% comparedto untreated or negative control treated subjects. In other embodiments,the change may be at least 100, 125, 150, 200, 250, 300, 350, 400, or450% compared to untreated or negative control treated subjects.

In another embodiment, the disclosure provides a method for decreasingamyloid plaque load in the brain of a subject. The method comprisesadministering a therapeutically effective amount of an anti-ApoEantibody to a subject. Suitable anti-ApoE antibodies include thosedisclosed herein. A method of the invention may decrease the amyloidplaque load in the hippocampus of a subject and/or decrease the amyloidplaque load in the brain cortex of a subject. In each of the aboveembodiments, the amyloid plaque load may be decreased by at least 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20%compared to untreated or negative control treated subjects. In someembodiments, the amyloid plaque load may be decreased by at least 20,25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95% comparedto untreated or negative control treated subjects. In other embodiments,the amyloid plaque load may be decreased by at least 100, 125, 150, 200,250, 300, 350, 400, or 450% compared to untreated or negative controltreated subjects.

In another embodiment, the disclosure provides a method for decreasingCAA load in the brain of a subject. The method comprises administering atherapeutically effective amount of an anti-ApoE antibody to a subjectwith fibrillar forms of Aβ in penetrating and/or leptomeningealarterioles on the surface of the cerebral cortex. Suitable anti-ApoEantibodies include those disclosed herein. A method of the invention maydecrease CAA load in the penetrating and/or leptomeningeal arterioles onthe surface of the cerebral cortex of a subject. In each of the aboveembodiments, CAA load may be decreased by at least 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20% compared tountreated or negative control treated subjects. In some embodiments, theamyloid plaque load may be decreased by at least 20, 25, 30, 35, 40, 45,50, 55, 60, 65, 70, 75, 80, 85, 90, or 95% compared to untreated ornegative control treated subjects. In other embodiments, the amyloidplaque load may be decreased by at least 100, 125, 150, 200, 250, 300,350, 400, or 450% compared to untreated or negative control treatedsubjects.

Another embodiment includes a method of reducing insoluble Aβ₄₂ levelsin the brain of a subject in need thereof. The method comprisesadministering a therapeutically effective amount of an anti-ApoEantibody to the subject. In one example, the method further comprisesreducing insoluble Aβ₄₀ levels in the brain of the subject. In anotherexample, the method comprises selectively reducing insoluble Aβ40levels, reducing insoluble Aβ42 levels, or a combination thereofcompared to soluble Aβ40, Aβ42 levels, or a combination thereof in thebrain of a subject.

The level of Aβ can be assessed by any suitable method known in the artcomprising, e.g., analyzing Aβ by one or more techniques chosen fromWestern blot, immunoprecipitation, enzyme-linked immunosorbent assay(ELISA), radioimmunoassay (RIA), fluorescent activated cell sorting(FACS), two-dimensional gel electrophoresis, mass spectroscopy (MS),matrix-assisted laser desorption/ionization-time of flight-MS(MALDI-TOF), surface-enhanced laser desorption ionization-time of flight(SEMI-TOE), high performance liquid Chromatography (HPLC), fast proteinliquid chromatography (FPLC), multidimensional liquid chromatography(LC) followed by tandem mass spectrometry (MS/MS), and laserdensitometry. In vivo imaging of Aβ is particularly suited forevaluating amyloid plaque load. Non-limiting examples of in vivo imagingmethods include positron emission tomography (PET), single photonemission tomography (SPECT), near infrared (NIR) optical imaging ormagnetic resonance imaging (MRI). Suitable imaging agents are also knownin the art (e.g. PIB.).

In another embodiment, the disclosure provides a method for improving anAβ plaque associated symptom and/or a CAA associated symptom in asubject. The method comprises administering a therapeutically effectiveamount of an anti-ApoE antibody that specifically binds ApoE to asubject with at least one Aβ plaque associated symptom and/or at leastone CAA associated symptom. Suitable anti-ApoE antibodies include thosedisclosed herein. Non-limiting examples of improved Aβ plaque associatedsymptoms are identified above. In certain embodiments, improved Aβplaque associated symptoms may include reduced neuronal degeneration,impaired cognitive function, altered behavior, emotional dysregulation,and/or seizures. In each of the above embodiments, the improvement (i.e.the change) in the symptom is at least statistically significant. Incertain embodiments, the change may be at least 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20% compared to untreatedor negative control treated subjects. In some embodiments, the changemay be at least 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85,90, or 95% compared to untreated or negative control treated subjects.In other embodiments, the change may be at least 100, 125, 150, 200,250, 300, 350, 400, or 450% compared to untreated or negative controltreated subjects.

For each of the above embodiments, suitable anti-ApoE antibodies aredescribed in Section II. In certain embodiments, an anti-ApoE antibodyis (a) selected from an antibody listed in Tables A-H, (b) is ananti-ApoE antibody that competitively inhibits HJ152 and/or HJ1514binding to human ApoE, or (c) is an anti-ApoE antibody thatcompetitively inhibits HJ155, HJ156, HJ159, HJ1513 and/or HJ1518 bindingto human ApoE. In certain embodiments, an anti-ApoE antibody is selectedfrom an antibody listed in Tables A-B, or is an anti-ApoE antibody thatcompetitively inhibits HJ152 and/or HJ1514 binding to human ApoE. Incertain embodiments, an anti-ApoE antibody is selected from an antibodylisted in Tables C-D, or is an anti-ApoE antibody that competitivelyinhibits HJ155, HJ156, HJ159, HJ1513 and/or HJ1518 binding to humanApoE. In certain embodiments, an anti-ApoE antibody is selected from anantibody listed in Tables E-F, or is an anti-ApoE antibody thatcompetitively inhibits HJ1531, and/or HJ1536 binding to human ApoE. Incertain embodiments, an anti-ApoE antibody is selected from an antibodylisted in Table G, or is an anti-ApoE antibody that competitivelyinhibits HJ151 binding to human ApoE. In certain embodiments, ananti-ApoE antibody is selected from an antibody listed in Table H, or isan anti-ApoE antibody that competitively inhibits HJ158 or HJ1526binding to human ApoE.

Anti-ApoE antibody disclosed herein can also be conjugated totherapeutic agents, prodrugs, peptides, proteins, enzymes, viruses,biological response modifiers, pharmaceutical agents, or PEG. In certainembodiments, therapeutic agent may be a drug, a radioisotope, a lectin,or a toxin. Conjugates that are immunotoxins have been widely describedin the art. The toxins can be coupled to the antibodies by conventionalcoupling techniques or immunotoxins containing protein toxin portionscan be produced as fusion proteins. In using radioisotopicallyconjugated anti-ApoE antibodies for immunotherapy, certain isotopes canbe chosen depending on such factors as leukocyte distribution as well asstability and emission. Depending on the autoimmune response, someemitters can be used. In general, a and P3 particle emittingradioisotopes are utilized in immunotherapy. In certain embodiments,short range, high energy a emitters such as ²¹²Bi can be used. Examplesof radioisotopes which can be bound to the anti-ApoE antibodiesdisclosed herein for therapeutic purposes include, but are not limitedto ¹²³I, ¹²⁴I, ¹²⁵I, ¹³¹I, ⁸⁹Zr, ⁹⁰Y, ⁶⁷Cu, ⁶⁴Cu, ¹¹¹In, ²¹²Bi, ²¹²At,²¹¹Pb, ⁴⁷Sc, ¹⁰⁹Pd, and ¹⁸⁸Re. Other therapeutic agents which can becoupled to the anti-ApoE antibodies, as well as ex vivo and in vivotherapeutic protocols, are known, or can be easily ascertained, by thoseof ordinary skill in the art.

Administration of an anti-ApoE antibody, or a composition comprising ananti-ApoE antibody, is performed using standard effective techniques,include peripherally (i.e. not by administration into the centralnervous system) or locally to the central nervous system. Peripheraladministration includes but is not limited to intravenous,intraperitoneal, subcutaneous, pulmonary, transdermal, intramuscular,intranasal, buccal, sublingual, or suppository administration. Localadministration, including directly into the central nervous system (CNS)includes but is not limited to via a lumbar, intraventricular orintraparenchymal catheter or using a surgically implanted controlledrelease formulation.

Pharmaceutical compositions for effective administration aredeliberately designed to be appropriate for the selected mode ofadministration, and pharmaceutically acceptable excipients such ascompatible dispersing agents, buffers, surfactants, preservatives,solubilizing agents, isotonicity agents, stabilizing agents and the likeare used as appropriate. Remington's Pharmaceutical Sciences, MackPublishing Co., Easton Pa., 16 Ed ISBN: 0-912734-04-3, latest edition,incorporated herein by reference in its entirety, provides a compendiumof formulation techniques as are generally known to practitioners. Itmay be particularly useful to alter the solubility characteristics ofthe antibodies useful in this discovery, making them more lipophilic,for example, by encapsulating them in liposomes or by blocking polargroups.

The concentration of antibody in formulations to be administered is aneffective amount and ranges from as low as about 0.1% by weight to asmuch as about 15 or about 20% by weight and will be selected primarilybased on fluid volumes, viscosities, and so forth, in accordance withthe particular mode of administration selected if desired. A typicalcomposition for injection to a living subject could be made up tocontain 1 mL sterile buffered water of phosphate buffered saline andabout 1-1000 mg of any one of or a combination of the antibodiesdisclosed herein. The formulation could be sterile filtered after makingthe formulation, or otherwise made microbiologically acceptable. Atypical composition for intravenous infusion could have volumes between1-250 mL of fluid, such as sterile Ringer's solution, and 1-100 mg perml, or more in anti-ApoE antibody concentration. Anti-ApoE antibodiesdisclosed herein can be frozen or lyophilized for storage andreconstituted in a suitable sterile carrier prior to use. Lyophilizationand reconstitution may lead to varying degrees of antibody activity loss(e.g. with conventional immune globulins, IgM antibodies tend to havegreater activity loss than IgG antibodies). Dosages administered areeffective dosages and may have to be adjusted to compensate. The pH ofthe formulations generally pharmaceutical grade quality, will beselected to balance antibody stability (chemical and physical) andcomfort to the subject when administered. Generally, a pH between 4 and8 is tolerated. Doses will vary from individual to individual based onsize, weight, and other physiobiological characteristics of theindividual receiving the successful administration.

As used herein, the term “therapeutically effective amount” means anamount of a substance (e.g. an anti-ApoE antibody) that leads tomeasurable and beneficial effects for the subject administered thesubstance, i.e., significant efficacy. The therapeutically effectiveamount or dose of compound administered according to this discovery willbe determined using standard clinical techniques and may be byinfluenced by the circumstances surrounding the case, including theantibody administered, the route of administration, and the status ofthe symptoms being treated, among other considerations. A typical dosemay contain from about 0.01 mg/kg to about 100 mg/kg of an anti-ApoEantibody described herein. Doses can range from about 0.05 mg/kg toabout 50 mg/kg, more preferably from about 0.1 mg/kg to about 25 mg/kg.The frequency of dosing may be daily or once, twice, three times or moreper week or per month, as needed as to effectively treat the symptoms.

The timing of administration of the treatment relative to the diseaseitself and duration of treatment will be determined by the circumstancessurrounding the case. Duration of treatment could range from a singledose administered on a one-time basis to a life-long course oftherapeutic treatments.

Although the foregoing methods appear the most convenient and mostappropriate and effective for administration of proteins such ashumanized antibodies, by suitable adaptation, other effective techniquesfor administration, such as intraventricular administration, transdermaladministration and oral administration may be employed provided properformulation is utilized herein. In addition, a person skilled in the artcan use a polynucleotide of the invention encoding any one of theabove-described antibodies instead of the proteinaceous material itself.For example,

In addition, it may be desirable to employ controlled releaseformulations using biodegradable films and matrices, or osmoticmini-pumps, or delivery systems based on dextran beads, alginate, orcollagen.

IV. Diagnosing or Tracking Methods

The present disclosure also provides anti-ApoE antibodies conjugated toa detectable signal (i.e. a measurable substance, or a substance thatgenerates a measurable signal). Non-limiting examples include variousenzymes, prosthetic groups, fluorescent materials, luminescentmaterials, bioluminescent materials, radioactive materials, positronemitting metals using various positron emission tomographies, andnonradioactive paramagnetic metal ions. See, e.g., U.S. Pat. No.4,741,900 for metal ions which can be conjugated to antibodies for useas diagnostics according to the disclosure. Examples of suitable enzymesinclude horseradish peroxidase, alkaline phosphatase, 3-galactosidase,or acetylcholinesterase; examples of suitable prosthetic group complexesinclude streptavidin/biotin and avidin/biotin; examples of suitablefluorescent materials include umbelliferone, fluorescein, fluoresceinisothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansylchloride or phycoerythrin; an example of a luminescent material includesluminol; examples of bioluminescent materials include luciferase,luciferin, and aequorin; and examples of suitable radioactive materialinclude ¹²⁵I, ¹³¹I, ¹¹¹In or ⁹⁹Tc. The signal generated by the agent canbe measured, for example, by single-photon emission computed tomography(SPECT) or positron emission tomography (PET).

Anti-ApoE antibodies conjugated to a detectable signal may be useddiagnostically to, for example, monitor the development or progressionof a neurodegenerative disease as part of a clinical testing procedureto, e.g., determine the efficacy of a given treatment and/or preventionregimen. The patient's treatment can be adjusted based on the level ofneurodegenerative disease progression.

In some embodiments, a method of assessing disease progression in asubject being treated for a neurodegenerative disease characterized bybrain amyloid plaques or a neurodegenerative disease characterized byvascular amyloid plaques in the brain is provided, the methodcomprising: (a) administering an anti-ApoE antibody disclosed hereinthat is labeled with an agent that generates a measurable signal asdescribed herein (i.e. “labeled anti-ApoE antibody”), wherein the signalis measured in the patient following the administration; (b)administering the labeled anti-ApoE or antigen-binding fragment thereofat one or more time intervals following the administration of (a),wherein the signal is measured in the patient following theadministration; and (c) assessing disease progression in the patientbased on a change in the signal measured in the patient at the one ormore time intervals following administration of (a); wherein an increasein the signal indicates progression of the neurodegenerative disease inthe patient. In certain embodiments, the subject is being treated withthe same anti-ApoE antibody, but in an unlabeled form. In certainembodiments, the subject is being treated with an anti-ApoE antibodythat competitively inhibits the labeled anti-ApoE antibody binding tohuman ApoE. In certain embodiments, the subject is being treated with ananti-ApoE antibody that does not competitively inhibit the labeledanti-ApoE antibody binding to human ApoE. In certain embodiments, thesubject is being treated with other drugs known in the art.

In other embodiments, a method of assessing disease progression in asubject being treated for a neurodegenerative disease characterized bybrain amyloid plaques or a neurodegenerative disease characterized byvascular amyloid plaques in the brain is provided, the methodcomprising: (a) administering an anti-ApoE antibody disclosed hereinthat is labeled with an agent that generates a measurable signal asdescribed herein (i.e. “labeled anti-ApoE antibody”), wherein the signalis measured in the patient following the administration; (b) assessingthe disease state in the subject upon review of a comparison of thesignal measured in the subject to the signal measured followingadministration of the labeled antibody or antigen-binding fragmentthereof to one or more control subjects; wherein an increase in thesignal generated in the patient relative to the control subjectcorrelates with an increase in brain amyloid plaques; and (c) treatingthe patient with a therapy appropriate for the patient's disease state.A “control subject(s),” refers to any normal healthy subject (or a poolof subjects), a subject or subjects with different degrees of disease,or even the actual test subject at an earlier stage of disease. Incertain embodiments, the therapy is the same anti-ApoE antibody, but inan unlabeled form. In certain embodiments, the therapy is an anti-ApoEantibody that competitively inhibits the labeled anti-ApoE antibodybinding to human ApoE. In certain embodiments, the therapy is ananti-ApoE antibody that does not competitively inhibit the labeledanti-ApoE antibody binding to human ApoE. In certain embodiments, thetherapy is with an anti-Aβ antibody, an anti-tau antibody, agamma-secretase inhibitor, a beta-secretase inhibitor, a cholinesteraseinhibitor, an NMDA receptor antagonist, or other drugs known in the art.

The present disclosure also provides the use of the anti-ApoE antibodiesdisclosed herein for measuring the amount of brain amyloid plaques in atest subject, assessing disease progression in a patient being treatedfor a neurodegenerative disease or treating a neurodegenerative diseasecharacterized by brain amyloid plaques in a patient in need oftreatment.

V. Pharmaceutical Compositions

The present disclosure encompasses pharmaceutical compositionscomprising an anti-ApoE antibody disclosed in Section II, so as tofacilitate administration and promote stability of the active agent. Forexample, an anti-ApoE antibody of this disclosure may be admixed with atleast one pharmaceutically acceptable carrier or excipient resulting ina pharmaceutical composition which is capably and effectivelyadministered (given) to a living subject, such as to a suitable subject(i.e. “a subject in need of treatment” or “a subject in need thereof”).Methods of preparing and administering anti-ApoE antibodies disclosedherein to a subject in need thereof are well known to or are readilydetermined by those skilled in the art. The route of administration ofan anti-ApoE antibody can be, for example, peripheral, oral, parenteral,by inhalation or topical.

Pharmaceutical compositions for effective administration aredeliberately designed to be appropriate for the selected mode ofadministration, and pharmaceutically acceptable excipients such ascompatible carriers, dispersing agents, buffers, surfactants,preservatives, solubilizing agents, isotonicity agents, stabilizingagents and the like are used as appropriate.

Non-limiting examples of pharmaceutically acceptable carriers, includephysiological saline, ion exchangers, alumina, aluminum stearate,lecithin, serum proteins, such as human serum albumin, buffer substancessuch as phosphates, glycine, sorbic acid, potassium sorbate, partialglyceride mixtures of saturated vegetable fatty acids, water, salts orelectrolytes, such as protamine sulfate, disodium hydrogen phosphate,potassium hydrogen phosphate, sodium chloride, zinc salts, colloidalsilica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-basedsubstances, polyethylene glycol, sodium carboxymethylcellulose,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers,polyethylene glycol, wool fat or a combination thereof.

Prevention of the action of microorganisms can be achieved by variousantibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, ascorbic acid, thimerosal and the like. In manycases, isotonic agents can be included, for example, sugars,polyalcohols, such as mannitol, sorbitol, or sodium chloride in thecomposition.

Prolonged absorption of the injectable compositions can be brought aboutby including in the composition an agent which delays absorption, forexample, aluminum monostearate and gelatin.

Compositions disclosed herein can be frozen or lyophilized for storageand reconstituted in a suitable sterile carrier prior to use.

In some embodiments, anti-ApoE antibodies may be formulated forparenteral administration. Preparations for parenteral administrationinclude sterile aqueous or non-aqueous solutions, suspensions, andemulsions. Examples of non-aqueous solvents are propylene glycol,polyethylene glycol, vegetable oils such as olive oil, and injectableorganic esters such as ethyl oleate. Aqueous carriers include water,alcoholic/aqueous solutions, emulsions or suspensions, including salineand buffered media. Parenteral vehicles include sodium chloridesolution, Ringer's dextrose, dextrose and sodium chloride, lactatedRinger's, or fixed oils. Intravenous vehicles include fluid and nutrientreplenishers, electrolyte replenishers (such as those based on Ringer'sdextrose), and the like. Preservatives and other additives can also bepresent such as, for example, antimicrobials, anti-oxidants, chelatingagents, and inert gases and the like. Parenteral formulations can be asingle bolus dose, an infusion or a loading bolus dose followed with amaintenance dose. These compositions can be administered at specificfixed or variable intervals, e.g., once a day, or on an “as needed”basis.

Certain pharmaceutical compositions, as disclosed herein, can be orallyadministered in an acceptable dosage form including, e.g., capsules,tablets, aqueous suspensions or solutions. Certain pharmaceuticalcompositions also can be administered by nasal aerosol or inhalation.Such compositions can be prepared as solutions in saline, employingbenzyl alcohol or other suitable preservatives, absorption promoters toenhance bioavailability, and/or other conventional solubilizing ordispersing agents.

The amount of an anti-ApoE antibody to be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. The compositioncan be administered as a single dose, multiple doses or over anestablished period of time in an infusion. Dosage regimens also can beadjusted to provide the optimum desired response (e.g., a therapeutic orprophylactic response).

The following examples are included to demonstrate preferred embodimentsof the invention. It should be appreciated by those of skill in the artthat the techniques disclosed in the examples that follow representtechniques discovered by the inventors to function well in the practiceof the invention. Those of skill in the art should, however, in light ofthe present disclosure, appreciate that many changes can be made in thespecific embodiments that are disclosed and still obtain a like orsimilar result without departing from the spirit and scope of theinvention, therefore all matter set forth or shown in the accompanyingexamples and drawings is to be interpreted as illustrative and not in alimiting sense

EXAMPLES

The following examples illustrate various iterations of the invention.

Example 1

Murine monoclonal antibodies to ApoE were generated and sequenced.Briefly, to generate the antibodies, human recombinant ApoE4 protein(299 amino acids) was injected intraperitoneally (IP) into a wildtypemouse on a B6C3 background. 100 μg of antigen (in 200 μl PBS+200 μlcomplete Freund's adjuvant) was injected on day 0, day 14 and day 28. Alast boost of 50 μg of antigen in PBS was injected IP 3 days beforefusion of myeloma cells with spleen cells of the mice. Serum was testedby direct ELISA to ApoE4 on day 21 and day 35. If titer was over1:10,000, myeloma cells were then fused with mouse spleen cells perstandard protocol, followed by isolation of hybridoma clones.

TABLE 1 Light chain Heavy chain Antibody variable region variable regionHJ151 SEQ ID NO: 1 SEQ ID NO: 2 HJ152 SEQ ID NO: 3 SEQ ID NO: 4 HJ153SEQ ID NO: 5 SEQ ID NO: 6 HJ154 SEQ ID NO: 7 SEQ ID NO: 8 HJ155 SEQ IDNO: 9 SEQ ID NO: 10 HJ156 SEQ ID NO: 11 SEQ ID NO: 12 HJ158 SEQ ID NO:97 SEQ ID NO: 98 HJ159 SEQ ID NO: 13 SEQ ID NO: 14 HJ1513 SEQ ID NO: 15SEQ ID NO: 16 HJ1514 SEQ ID NO: 17 SEQ ID NO: 18 HJ1518 SEQ ID NO: 19SEQ ID NO: 20 HJ1526 SEQ ID NO: 21 SEQ ID NO: 22 HJ1531 SEQ ID NO: 115SEQ ID NO: 116 HJ1536 SEQ ID NO: 121 SEQ ID NO: 122

Example 2

Brain lysates from apoE KO mice or mice expressing human apoE3, humanapoE4 or murine apoE were immunoblotted with HJ151, HJ152, HJ153, andHJ154. A commercial apoE antibody GA-50 was used as a positive control.FIG. 1 and FIG. 2 show that, by Western blot, HJ152, HJ153, and HJ154recognize both ApoE3 and ApoE4, and HJ151 is ApoE4 specific.

Example 3

Brain tissue from 5XFAD APP transgenic mice expressing different humanApoE isoforms were stained using biotinylated HJ153 and HJ154. Braintissue from ApoE KO mouse was used as negative control. As shown in FIG.3, HJ153 and HJ154 stain ApoE in the neuropil, in astrocytes, and inamyloid plaques, if plaques are present.

Example 4

Phosphate buffered saline brain cortical tissue lysates containingApoE2, ApoE3 or ApoE4 were immunoprecipitated using HJ152, HJ153 andHJ154. Materials immunoprecipitated with the antibodies (labeled as IP)and the solution after immunoprecipitation were immunoblotted afterrunning an SDS-PAGE gel and transfer to nitrocellulose membrane usingcommercial apoE antibody GA-50. WUE4, a monoclonal antibody againsthuman apoE, was used as a positive control. As shown in FIG. 4, HJ152 isable to immunoprecipitate some of the apoE from the samples, thoughthere is still some apoE remaining in the sample afterimmunoprecipitation (compare IP vs. post IP, respectively). FIG. 4 alsoshows that HJ153 and HJ154 are able to immunoprecipitate all ApoE fromthe samples.

Example 5

ELISA plates were coated with 0.5 μg/ml of recombinant apoE2, apoE3, orapoE4. Then different concentrations of HJ151 (FIG. 5A), HJ153 (FIG.5B), HJ154 (FIG. 5C), or HJ156 (FIG. 5D) were loaded on the plates.Horse-radish peroxidase labeled goat anti-mouse secondary antibodieswere used to detect binding. FIG. 5A-D shows that HJ153 and HJ154 bindApoE2, ApoE3 and ApoE4 as detected in the ELISA. HJ151 is specific forApoE4 and HJ156 binds only ApoE3 and ApoE4. Surface plasmon resonanceprofiles for various antibodies was also performed. Anti-ApoE antibodieswere serially diluted 3-fold (starting at 100 nM for HJ153 (FIG. 5F),and 1000 nM for HJ151 (FIG. 5E), and HJ156 (FIG. 5G) for detection ofbinding to biotinylated-recombinant apoE4 captured on a streptavidinchip. Samples were injected at a flow rate of 30 μl/minute. FIG. 5Hshows apparent KD values of HJ151, HJ153 and HJ156 which were calculatedbased on the SPR experiment.

Example 6

APP/PS1-21 E4/E4 mice received continuous intracerebroventricular (ICV)infusion of PBS (negative control), mouse IgG2 (negative control), HJ5.1(anti-Aβ antibody, positive control), or an anti-apoE antibody (HJ151,HJ154, or HJ156), beginning at 2 months of age (n=10-12/group).Anti-ApoE antibody or control antibody (2 mg/ml) was filled into asubcutaneous osmotic minipump (Alzet, model 2006) and infused through asurgically implanted catheter into the left lateral cerebral ventricle(bregma −0.4 mm, 1.0 mm lateral to midline, 2.5 mm below the skull),infusing fluid at the speed of 1.2 μl/min for 6 weeks.

At the age of 3.5-month, the mice were perfused and brain sections werestained for either Aβ plaques using anti-Aβ antibody HJ3.4B or fibrillarplaques using Thioflavine S. The percent of area covered by plaques inthe cerebral cortex dorsal to hippocampus was quantified. One-way ANOVAfollowed by Tukey post-test was used to analyze the data. As shown inFIG. 6 and FIG. 7, there was a statistically significant decrease in thepercent area covered by Aβ and fibrillar plaques following ICV infusionof certain anti-ApoE antibodies. The amount of soluble and insoluble Aβwas also quantified in the cerebral cortex dorsal to hippocampus byELISA. Briefly, cortical tissue was homogenized in PBS, Triton, and 5Mguanidine sequentially, and the Aβ levels in each fraction weremeasured. A statistically significant decrease in insoluble Aβ42 (Tritonand guanidine fractions) was observed for HJ151 and HJ156 (FIG. 8).

Example 7

APP/PS1-21 E4/E4 mice received an intraperitoneal injection of PBS(negative control), mouse IgG (negative control), or an anti-apoEantibody (HJ151, HJ155, and HJ156) beginning at 2 months of age(n=10/group). Antibodies were injected once per week at 50 mg/kg. Bodyweight of each mouse was measured at the time of injection. As shown inFIG. 9, there was no difference in body weight between treatment groups.Three days after the seventh injection, the mice were perfused. The lefthemi-brain was fixed in 4% PFA at 4° C. for 48 hrs, then transferred toPBS and stored at 4° C. The right hemi-brain was dissected and frozenfor biochemical analysis.

The amount of soluble and insoluble Aβ was quantified in the cerebralcortex dorsal to hippocampus by ELISA. Briefly, cortical tissue washomogenized in PBS and 5M guanidine sequentially, and the Aβ levels insoluble (PBS) and insoluble (guanidine) fractions were measured. Asshown in FIG. 10-13, HJ156 significantly decreased insoluble Aβ40 andAβ42 (p<0.0001) but did not have a statistically significant effect onsoluble Aβ. The level of insoluble Aβ40 and 42 were also decreased byHJ155 and HJ151 but the decrease was not significant.

Example 8

APP/PS1-21 E4/E4 mice were injected with an AAV2/8 vector producinganti-ApoE antibodies (HJ151, HJ151Δ, or HJ156) or controls (HJ16.5 andPBS) (n=24/group). The AAV serotype was AAV2/8 and the promoter was achicken beta actin (CBA) promoter. The injections were done withbilateral intracerebroventricular injections in PO mice. The HJ151Δantibody is the HJ151 antibody with a mutation in the Fc domain, whichprevents binding to Fc gamma receptors. The antibody HJ16.5 binds to thehuman protein PLD3 and does not bind to murine PLD3 or other mouseproteins. At the age of 3.5 months, the mice were perfused. The lefthemi-brain was fixed in 4% PFA at 4° C. for 48 hrs, then transferred to30% sucrose and stored at 4° C. The right hemi-brain was dissected andfrozen for biochemical analysis.

The amount of soluble and insoluble Aβ was quantified in the cerebralcortex dorsal to hippocampus by ELISA. Briefly, cortical tissue washomogenized in PBS and 5M guanidine sequentially, and the Aβ levels insoluble (PBS) and insoluble (guanidine) fractions were measured. Asshown in FIGS. 15 and 16, antibodies HJ151 and HJ156 significantlydecreased insoluble Aβ40 and Aβ42, whereas HJ151Δ and HJ156Δ had noeffect. None of the antibodies had a significant effect of soluble Aβ40or Aβ42 (not shown). As shown in FIG. 14, relative antibodyconcentrations in the cortex of APPPS1-21/APOE4 mice expressingrecombinant (r) HJ151, r HJ151 with D265A mutation (Δ), rHJ156, andrHJ156Δ were determined by antigen capture ELISA. The relative level ofeach antibody was calculated by using its hybridoma-derived, purifiedantibody as a standard. Similar levels of expression were found and nostatistical differences between the different antibodies.

Overall, expression of HJ151 and HJ156 by AAV significantly reducedinsoluble Aβ40 and Aβ42 in the APP/PS1-21 E4/E4 mouse model. Mutation ofFc effector (D265A) completely abolished the ability of HJ151 to reduceinsoluble Aβ. These results indicate clearance of Aβ aggregates byanti-apoE antibody may require an antibody opsonization and phagocytosisstep involving microglia mediated Aβ clearance.

Example 9

Unfixed tissue sections (20 μm thickness) from APP/PS1-21 E4/E4(APPPS1-21/E4), 5XFAD/apoE knockout (5XFAD/EKO) mice and humanapoE^(4/4) (human E4) brains from persons who died with amyloiddeposition were stained using the biotinylated antibodies HJ3.4B(anti-Aβ), HJ151B, HJ152B, HJ153B, HJ154B, HJ155B, and HJ156B (anti-apoEantibodies). (Note: “B” indicates “biotinylated”.) All mice utilizedwere old enough to have a significant amount of Aβ in the cortex(APPPS1-21/apoE^(4/4), >6 months old; 5XFAD/apoE knockout(5XFAD/EKO), >6 months old). All images in the mice and human sectionsare from the cortex. The results shown in FIG. 18 demonstrate thatHJ3.4B, HJ151B, HJ152B, HJ153B, HJ154B, HJ155B, and HJ156B stain plaquesand cerebral amyloid angiopathy in the APP/PS1-21 E4/E4 and humanapoE^(4/4) brains. No staining is observed in 5XFAD/EKO brains withanti-apoE specific antibodies.

Example 10

The affinity of several anti-ApoE antibodies for ApoE4 was determined bysurface plasmon resonance using a BIAcore T200. Anti-biotin monoclonalmouse antibody was immobilized on a Biacore Series S CM5 sensor chip.Biotinylated recombinant alipidated ApoE4 was then captured on to thechip. Serial dilutions of each antibody were injected at a flow rate of30 μl/min. Each sample was analyzed, for example, with 3-minuteassociation and 10-minute dissociation. After each injection the chipwas regenerated using 3 M MgCl₂ or another appropriate buffer. Bindingresponse was corrected by subtracting the response units (RUs) from aflow cell capturing an irrelevant IgG at similar density. A 1:1 Languirmodel of simultaneous fitting of k_(on) and k_(off) was used forkinetics analysis. The data are shown in Table 2. Recombinant alipidatedApoE4 was either expressed in soluble form in E. coli and then purified,or purchased from a commercial supplier.

TABLE 2 Antibody k_(on) (1/Ms) k_(off) (1/s) KD (M) Rmax (RU) HJ1511.97E+04 1.19E−03 6.02E−08 77.5 HJ152 4.66E+03 1.16E−03 2.49E−07 176HJ153 8.04E+05 3.46E−04 4.30E−10 261.9 HJ154 5.16E+05 3.48E−04 6.75E−10262.9 HJ156 2.07E+04 4.49E−03 2.16E−07 78 HJ159 1.89E+04 7.40E−033.91E−07 71.7 HJ1513 6.36E+03 8.58E−04 1.35E−07 226.5 HJ1514 1.28E+04<1.00E−05  <7.81E−10  43.5 HJ1518 4.59E+04 9.46E−03 2.06E−07 56 HJ15267.88E+03 5.28E−03 6.70E−07 34.7

Example 11

ELISA dose response curves were generated for several anti-ApoEantibodies. Briefly, plates were coated overnight with full lengthrecombinant alipidated ApoE2, ApoE3, or ApoE4 (0.5 μg/ml). The followingdays, plates were washed, blocked and then incubated with an anti-ApoEantibody. A goat, anti-mouse IgG-HRP antibody was used for secondarydetection. The data are shown in Table 3. Recombinant alipidated ApoEwas either expressed solubly in E. coli and then purified, or purchasedfrom a commercial supplier.

TABLE 3 Binding EC50s (nM) against ApoE isoforms IC 50_E2 IC 50_E3 IC50_E4 HJ151 >1000 >1000 0.44 HJ152 >1000 0.13 0.35 HJ153 0.02 0.02 0.03HJ154 0.25 0.24 0.26 HJ155 >1000 0.78 1.1 HJ156 >1000 1.69 0.62 HJ1580.07 0.09 0.06 HJ159 >1000 1.90 0.32 HJ1513 ~100 1.26 0.46 HJ1514 ~1000.23 0.04 HJ1518 >1000 0.258 0.06 HJ1526 0.03 0.05 0.04 HJ1531 0.05 0.050.04 HJ1536 0.09 0.10 0.05

Example 12

Anti-ApoE antibodies were assessed for detection of ApoE derived fromhuman plasma lipoprotein by a coated plasma ELISA (FIG. 19A). Humanplasma (identified as possessing one or more e4 allele by western blotassay with HJ151 and comprising about 1-5 μM ApoE4) was diluted 100× inPBS and coated onto half-well ELISA microtiter plates overnight at 4° C.Plates were washed extensively in PBS and blocked with 5% BSA/PBS for 1hour. Antibodies were added to the wells at 400-500 nM startingconcentration with 5-fold dilutions thereafter for 1 hour. Binding ofantibodies was detected by an anti-mouse IgG secondary conjugated tohorse-radish peroxidase (HRP). TMB was the detection substrate with 4Nsulfuric acid as the stop solution.

Compared to HJ153 and HJ154, HJ156 demonstrated reduced binding tocoated human plasma ApoE (FIG. 19B). A control IgG and a commerciallyavailable anti-ApoE antibody, WUE4, showed minimal binding. These datasuggest HJ153 and HJ154 robustly detect lipidated ApoE contained inlipoprotein particles from human plasma. FIGS. 19C and D show thebinding profiles of other antibodies for coated human plasma. HJ159,HJ1514, HJ1518, HJ1526 (FIG. 19C), HJ158, HJ1531, and HJ1536 (FIG. 19D)did not detect lipidated, plasma ApoE robustly at the highest antibodyconcentration and minimally at the next dilution downward.

Example 13

Antibodies that failed to detect immobilized human ApoE from plasma werefurther tested in a competition ELISA to rule out the possibility thatcoated plasma ApoE epitopes were unavailable for binding due tointeraction with the plastic wells (FIG. 20A). Recombinant alipidatedApoE was coated onto half-well ELISA microtiter plates at 0.5 ug/mL inPBS overnight at 4° C. Plates were washed extensively in PBS and blockedwith 5% BSA/PBS for 1 hour. HJ153 and antibody clones that did notdetect coated human plasma ApoE were pre-incubated in a dilution seriesof human plasma (comprising about 1-5 μM ApoE4)/5% BSA/PBS (startingdilution 5×, 2-fold thereafter) and either 4 nM or 50 nM of antibody.After 1 hour of pre-incubation, the mixture was added to the blockedplate and incubated for 1 hour. Detection of antibody available forbinding to coated alipidated recombinant ApoE4 (not competed off by ApoEin diluted plasma) was with anti-mouse-IgG-HRP and TMB. The reaction wasstopped with 4N sulfuric acid.

The competition ELISA showed titration of 4 nM HJ153 for binding tocoated ApoE4 (FIG. 20B) with increasing plasma dilutions. Antibodiesthat did not bind plasma in the previous assay did not show titration ofbinding to coated recombinant alipidated ApoE4 at either 4 nM or 50 nMantibody concentrations. These results suggest that antibodies that donot detect lipidated ApoE in plasma may prefer to bind recombinantalipidated ApoE.

Example 14

Unfixed tissue sections from APP/PS1-21 E2/E2, APP/PS1-21 E3/E3, andAPP/PS1-21 E4/E4 mice were stained for ApoE using the biotinylatedantibody HJ156 (“HJ156B”). Mice were 9 months old at the time ofstaining. Section thickness was 20 microns. All mice were perfusedtranscardially with PBS at 4 degrees C. prior to removal of the brain.The results are shown in FIG. 21-23. All images are from the cortex.

Example 15

Unfixed tissue sections from APP/PS1-21 E4/E4 mice were stained for ApoEusing anti-ApoE msIgG/huFc chimeric antibody HJ153, anti-ApoE msIgG/huFcchimeric antibody HJ156, anti-ApoE msIgG/huFc chimeric antibody HJ1514,or anti-ApoE msIgG/huFc chimeric antibody HJ1518. A biotinylated rabbitanti-human IgG antibody was used as the secondary antibody. Mice were 9months old at the time of staining. Section thickness was 20 microns.All mice were perfused transcardially with PBS at 4 degrees C. prior toremoval of the brain. The results are shown in FIG. 24-28. All imagesare from the cortex.

Example 16

APP/PS1-21 E4/E4 mice received intraperitoneal injections of PBS(negative control), HJ6.6 (IgG negative control), or an anti-apoEantibody (HJ151, HJ155, and HJ156) beginning at 2 months of age(n=10/group). Antibodies were injected once per week at 50 mg/kg until3.5 months of age. Mice per perfused transcardially with PBS at 4degrees C. prior to brain removal. Brains were fixed in 4%paraformaldehyde. Sections were cut at 50 microns thickness prior tostaining with the indicated reagents.

Brain sections from 2 month old untreated and 3.5 month old treated micewere immunostained with anti-Aβ antibody HJ3.4-biotin or stained withX-34 (a dye that stains fibrillar plaques), and staining was quantifiedfrom cortex. Treatment of APP/PS1-21 E4/E4 mice with HJ156 leads to astatistically significant reduction in Aβ plaque load, measured eitherby % area Aβ deposition (FIG. 29) or by plaque number (FIG. 30).Intra-peritoneal administration of HJ155 also resulted in astatistically significant decrease in Aβ plaque number (FIG. 30); adecrease in Aβ deposition (as measured by % area Aβ staining) was alsoobserved, though the decrease was not significant (FIG. 29). Similarreductions in fibrillar Aβ plaque deposition and number were observed byX-34 staining (FIGS. 31 and 32). Representative images from these brainsections are shown in FIG. 33-36.

Example 17

To investigate whether the ability to recognize conformationallydistinct forms of ApoE in Aβ plaques results in a more potent plaquereducing antibody, HJ151, HJ153 and HJ156 were further characterized.

HJ153 or HJ156 were coated onto plates and plasma from ApoE KO (EKO),APOE2, APOE3 and APOE4 mice was loaded (FIG. 37C and FIG. 37D). Theresults demonstrated that HJ153 bound to lipidated ApoE2, ApoE3 andApoE4 (FIG. 37C) while HJ156 did not (FIG. 37D). Next, non-lipidatedApoE4 was coated onto the plates and HJ151, HJ153 and HJ156pre-incubated with serially diluted plasma from APOE4 mice. The resultsshowed that lipidated ApoE was able to compete with the unlipidated ApoEfor HJ153, suggesting that HJ153 bound both lipidated and unlipidatedforms of ApoE (FIG. 37E). However, the lipidated ApoE was not able tocompete for the binding between non lipidated apoE with HJ151 or HJ156,suggesting that HJ151 and HJ156 preferentially bind to non lipidatedforms of ApoE (FIG. 37E).

Chimeric versions of HJ153 and HJ156 were also characterized. Plasmafrom EKO, APOE2, APOE3 and APOE4 KI mice were coated to ELISA plates(with plasma lipoprotein present). Chimeric versions of HJ153 (chi153)and HJ156 (chi156) that contain the binding domains of the mouse IgGantibodies with a human IgG backbone were prepared. Differentconcentrations of the chimeric antibodies were loaded onto the plates.The chi153 and chi156 that bound to the plate were detected usinganti-human IgG (FIG. 37A, FIG. 37B). The results reveal that chi153 wasable to bind lipidated ApoE2, ApoE3 and ApoE4 (FIG. 37A) while chi156did not (FIG. 37B).

The pharmacokinetic characteristics of HJ153 and HJ156 after IPinjection (10 mg/kg body weight) into APOE4 and APOEKO mice (FIG. 37F)were also compared. Plasma HJ153 was below detection limit at timepoints after 48 hr post-injection while HJ156 was present in the plasma14 days after injection (FIG. 37F). In summary, our ELISA data suggeststhat HJ156 preferentially binds to non lipidated ApoE while HJ153 bindsto both lipidated and non lipidated ApoE. This is consistent with alikely explanation for the pharmacokinetic characteristics. HJ153 bindsto lipidated ApoE which is abundant in the plasma, resulting in a higherclearance rate of HJ153. However, the fact that HJ156 only binds nonlipidated ApoE which is low or absent in plasma led to a slowerclearance of HJ156 in vivo.

In order to test whether HJ151 and HJ156 bind to some form of ApoE inthe brain, unfixed frozen brain sections from APPPS1/APOE4 or APPPS1/EKOmice using HJ151 and HJ156 (FIG. 38A). The presence of Aβ plaques wasconfirmed with Aβ immunostaining using anti-Aβ antibody HJ3.4 on thesections from the same brain. The results demonstrated that HJ151 andHJ156 were able to bind ApoE in plaques (FIG. 38A). Next, the ability ofHJ156 to bind to an aggregated form of ApoE induced by incubation at 40°C. was queried. The binding of untreated ApoE to HJ151, HJ153 or HJ156was compared to the binding of ApoE incubated at 40° C. to HJ151, HJ153or HJ156. HJ153 bound to untreated ApoE or ApoE incubated at 40° C.similarly while HJ151 and HJ156 preferentially bound to ApoE incubatedat 40° C. (FIG. 38B). It was further confirmed that 40° C. incubationresulted in the formation of aggregates in the pellet fraction afterultracentrifugation at 186,000×g for 1 hr (FIG. 38C). When the ApoE fromsupernatant or the pellets were coated onto ELISA plates and detectedwith HJ156, the ApoE from the supernatant showed low binding while theApoE in the pellet demonstrated high binding (FIG. 38D). The ApoE fromthe pellet or total incubated ApoE bound with HJ156 to a similar degree(FIG. 38D), suggesting that the HJ156 binding to ApoE species followingtreatment exposure to 40° C. are the ApoE species in the pellet.Solubilization of the ApoE aggregates reduced their binding to HJ156.Specifically, exposure of the ApoE aggregates to 1% SDS or 4M guanidinewas able to eliminate the binding of HJ156 to ApoE aggregates induced byincubation at 40° C. (FIG. 38E). In summary, the data showed that HJ156preferentially bound to aggregated forms of ApoE as compared tomonomeric ApoE. It also bound to ApoE in the plaques in APPPS1 mousebrain sections.

To test whether HJ151 and HJ156 bind to ApoE in the brain of livingmice, experiments were performed utilizing in vivo 2-photon microscopy.A control mouse IgG2 antibody, HJ151 and HJ156 were conjugated withAlexa 594 and applied to the surface of the brain (40 μl of 1 mg/ml) of6 month old APPPS1-21/APOE4 mice. The binding of the antibodies to ApoEwas monitored through a cranial window using 2-photon microscopy (FIG.39A-C). The results demonstrated HJ151 and HJ156 bind ApoE in theamyloid plaques and CAA. To determine whether peripherally administeredApoE antibodies can enter the brain and bind to ApoE, chi156 or controlhuman IgG were IP injected (50 mg/kg body weight) for 1 or 2 doses intothe APPPS1-21/APOE4 mice. Two days after the final injection, chi156 wasdetected bound to plaques in mice that received 1 or 2 injections of theantibody (FIG. 39D). In summary, the data demonstrated that HJ156 wasable to enter the brain and bind to ApoE in the Aβ plaques in livinganimals.

To determine whether HJ151 and HJ156 antibodies can increase activatedmicroglia, CD45+ microglia were quantified relative to the amount offibrillar plaques after short-term treatment of HJ151 and HJ156antibodies (4 doses of IP injection every 3 days) in APPPS1-21/APOE4mice that already had existing plaques at 4 months of age (FIG. 40A,FIG. 41). After acute passive immunization, HJ151 had no effects on theamount of activated microglia while HJ156 significantly increased theCD45+ microglia as compared to the controls (FIG. 40). Effects onmicroglial activation after acute peripheral injection may be related toantibody efficacy and dose since both of these antibodies can bind toApoE in plaques.

To determine the dose-range efficacy profile of HJ156, APPPS1-21/APOE4mice were IP injected with a weekly dose of HJ156 at one of three doselevels (2, 10 and 50 mg/kg) for 2.5 months starting at 2 months of age(mixed gender, n=17-18/group). Control groups received a weekly dose ofIgG2ab (50 mg/kg) or anti-Aβ HJ3.4 (50 mg/kg). Three days after thetwelfth injection, the mice were deeply anesthetized perfused. The righthemi-brain was dissected the amount of soluble and insoluble Aβ wasquantified in the cerebral cortex dorsal to hippocampus by ELISA.Cortical tissue was homogenized in PBS and 5M guanidine sequentially,and the Aβ levels in soluble (PBS) and insoluble (guanidine) fractionswere measured. There was a significant dose-dependent effect of HJ156 onreducing insoluble Aβ₄₀ and Aβ₄₂ levels in the brain (FIG. 42A-B).

Overall, the above data suggests that a key feature of the antibodiesdisclosed herein for decreasing Aβ plaques is the ability to selectivelybind a conformationally altered form of ApoE found in amyloid plaques.Given that non-lipidated ApoE is only a small percentage of total ApoEin vivo, targeting this pool of ApoE may be why no effect is seen onApoE levels or lipid metabolism. Moreover, it is likely that theseantibodies' preferential binding of alipidated ApoE contributes to alonger plasma half-life than as compared to an anti-ApoE antibody thatbinds to lipidated ApoE (FIG. 37F). This longer half-life would greatlyincrease the concentration of antibody that can enter the brain and bindto its target.

Example 18

Epitope binding of various anti-ApoE antibodies was evaluated multipleways. Different sequences of human ApoE were expressed on the surface ofyeast by yeast display using different plasmids. Different anti-ApoEantibodies were then assessed to determine binding to the different ApoEsequences by immunofluorescence of the yeast cell surface (Table 4).Because none of the anti-ApoE antibodies tested here recognize murineApoE, the epitope was further narrowed down by comparison of the murineand human ApoE sequences as shown in Tables 4 and 5. The data in Tables6 and 7 was generated as follows. DNA constructs containing differentregions of ApoE4 with myc tag were transformed into yeast cells. Yeastcells were incubated with HJ15x to detect binding or anti-myc antibodyto detect expression. HJ153 was not evaluated in these experiments as itwas found to not bind to human ApoE4 expressed in yeast.

TABLE 4 Antibody Specificity Epitope (aa of mature ApoE) 151 E4 108-120152 E2, E3, E4 150-160 153 E2, E3, E4  1-250 154 E2, E3, E4 130-140 155E2, E3, E4 150-160 156 E3, E4 140-150

TABLE 5 Antibody Epitope* 151 108-120 aa 152 150-160 aa 153 130-140 aa154 150-160 aa 155 140-150 aa 156 150-16 0aa *Numbering based off matureApoE protein (i.e., lacking the signal peptide).

TABLE 6 ApoE4 Expression amino in yeast acids Approximate region HJ151HJ152 HJ154 HJ155 HJ156 Yes  1-132 up to helix 3 + − + − − Yes  1-164 upto helix 4 + + + + + Yes  1-210 N terminal domain + + + + + Yes  1-299full length + + + + + Yes 24-79 helix1 + 2 − − − − − Yes  24-125helix1 + 2 + 3 + − + − − Yes  24-164 helix1 + 2 + 3 + 4 + + + + + Low 54-164 helix2 + 3 + 4 + + + + + No  88-164 helix3 + 4 No 132-164 LDLRbinding No 132-210 post helix 3 Yes 164-210 intermediate domain − − − −− Yes 210-299 C-terminal domain − − − − −

TABLE 7 ApoE4 Expression amino in yeast acids Approximate region HJ159HJ1513 HJ1514 HJ1518 HJ1526 Yes  1-132 up to helix 3 − − − − + Yes 1-164 up to helix 4 + + + + + Yes  1-210 N terminal domain + + + + +Yes  1-299 full length + + + + + Yes 24-79 helix1 + 2 − − − − − Yes 24-125 helix1 + 2 + 3 − − − − + Yes  24-164 helix1 + 2 + 3 +4 + + + + + Low  54-164 helix2 + 3 + 4 + + + + + No  88-164 helix3 + 4No 132-164 LDLR binding No 132-210 post helix 3 Yes 164-210 intermediatedomain − − − − − Yes 210-299 C-terminal domain − − − − −

Example 19

Clearance (CL) is a pharmacokinetic parameter that describes theefficiency of irreversible elimination of a drug from systemiccirculation, expressed as volume of blood/plasma/serum cleared of drugper unit time. Anti-ApoE antibodies that bind to lipidated ApoE in theplasma exhibit high clearance due to very high concentrations oflipidated ApoE present in circulation. In contrast, antibodies thatspecifically bind alipidated ApoE would be expected to exhibit lowclearance and other pharmacokinetic properties that are similar to anisotype control antibody that does not bind to antigen. For suchantibodies the clearance would not be dose-dependent.

Prior to the present disclosure, antibodies that have the ability topreferentially bind alipidated ApoE over lipidated ApoE were not known.The inventors have surprisingly found certain antibodies thatpreferentially bind alipidated ApoE over lipidated ApoE. Moreover,certain antibodies were found to bind with greater affinity toaggregated ApoE than to non-aggregated ApoE. The aggregated ApoE can beprepared by oxidation and/or heat treatment of ApoE (e.g., recombinantApoE). The aggregated ApoE can be separated from the non-aggregated fromby ultracentrifugation (at least 186,000 g).

Mice were administered antibody via intraperitoneal injections at theindicated doses and submandibular bleeds were performed at 30 minutes, 4hours, 1 day, 2 days, 4 days, and 7 days. Anti-ApoE drug levels weredetermined in the sampled plasma by an ELISA assay using recombinantapoE4 as a capture antigen for HJ156 or human Her2 protein for thecontrol IgG antibody (anti-Her2). Clearance values were calculated basedon overall pharmacokinetics for the antibodies over the entire timecourse of the experiment.

TABLE 8 Plasma antibody clearance values for HJ156 at various doses inAPOE4KI and APOEKO mice. Antibody Dose (mg/kg) Mouse CL (mL/day/kg)control mouse 10 ApoE4 knock-in 9.4 IgG HJ156 2 ApoE4 knock-in 20 HJ15610 ApoE4 knock-in 19 HJ156 50 ApoE4 knock-in 20 HJ156 10 ApoE4 knock-out13

1. An isolated anti-ApoE antibody comprising (a) a light chain variableregion comprising SEQ ID NO: 25 and optionally further comprising one ormore consensus sequences selected from SEQ ID NO: 78 or SEQ ID NO: 24,and/or (b) a heavy chain variable region comprising a consensus sequenceof SEQ ID NO: 84 and optionally further comprising one or more consensussequences selected from SEQ ID NO: 80 or SEQ ID NO:
 82. 2. The isolatedantibody of claim 1, wherein the consensus sequence of SEQ ID NO: 84 isSEQ ID NO:
 85. 3-4. (canceled)
 5. The isolated antibody of claim 1,wherein the light chain variable region comprises SEQ ID NO: 25 and oneor more consensus sequences selected from SEQ ID NO: 79 or SEQ ID NO:24; and/or the heavy chain variable region comprises a consensussequence of SEQ ID NO: 84 and one or more consensus sequences selectedfrom SEQ ID NO: 81 or SEQ ID NO:
 83. 6. (canceled)
 7. The isolatedantibody of claim 1, wherein the amino acid sequence of the light chainvariable region is SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ IDNO: 15, or SEQ ID NO: 19; or the amino acid sequence of the heavy chainvariable region is SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ IDNO: 16, or SEQ ID NO:
 20. 8. (canceled)
 9. The isolated antibody ofclaim 7, wherein (a) the amino acid sequence of the light chain variableregion is SEQ ID NO: 9 and the amino acid sequence of the heavy chainvariable region is SEQ ID NO: 10; (b) the amino acid sequence of thelight chain variable region is SEQ ID NO: 11 and the amino acid sequenceof the heavy chain variable region is SEQ ID NO: 12; (c) the amino acidsequence of the light chain variable region is SEQ ID NO: 13 and theamino acid sequence of the heavy chain variable region is SEQ ID NO: 14;(d) the amino acid sequence of the light chain variable region is SEQ IDNO: 15 and the amino acid sequence of the heavy chain variable region isSEQ ID NO: 16; or (e) the amino acid sequence of the light chainvariable region is SEQ ID NO: 19 and the amino acid sequence of theheavy chain variable region is SEQ ID NO:
 20. 10. The isolated antibodyof claim 1, wherein the consensus sequence of SEQ ID NO: 84 is SEQ IDNO:
 50. 11. The isolated antibody of claim 10, wherein the light chainvariable region further comprises one or more sequences selected fromSEQ ID NO: 47 or SEQ ID NO: 24; and/or the heavy chain variable regionfurther comprises one or more sequence selected from SEQ ID NO: 48 orSEQ ID NO:
 49. 12. (canceled)
 13. The isolated antibody of claim 1,wherein the consensus sequence of SEQ ID NO: 84 is SEQ ID NO:
 54. 14.The isolated antibody of claim 13, wherein the light chain variableregion further comprises one or more sequences selected from SEQ ID NO:51 or SEQ ID NO: 24; and/or the heavy chain variable region furthercomprises one or more sequence selected from SEQ ID NO: 52 or SEQ ID NO:53.
 15. (canceled)
 16. The isolated antibody of claim 1, wherein theconsensus sequence of SEQ ID NO: 84 is SEQ ID NO:
 58. 17. The isolatedantibody of claim 16, wherein the light chain variable region furthercomprises one or more sequences selected from SEQ ID NO: 55 or SEQ IDNO: 24; and/or the heavy chain variable region further comprises one ormore sequence selected from SEQ ID NO: 56 or SEQ ID NO:
 57. 18.(canceled)
 19. The isolated antibody of claim 1, wherein the consensussequence of SEQ ID NO: 84 is SEQ ID NO:
 62. 20. The isolated antibody ofclaim 19, wherein the light chain variable region further comprises oneor more sequences selected from SEQ ID NO: 59 or SEQ ID NO: 24; and/orthe heavy chain variable region further comprises one or more sequenceselected from SEQ ID NO: 60 or SEQ ID NO:
 61. 21. (canceled)
 22. Theisolated antibody of claim 1, wherein the consensus sequence of SEQ IDNO: 84 is SEQ ID NO:
 71. 23. The isolated antibody of claim 22, whereinthe light chain variable region further comprises one or more sequencesselected from SEQ ID NO: 68 or SEQ ID NO: 24; and/or the heavy chainvariable region further comprises one or more sequence selected from SEQID NO: 69 or SEQ ID NO:
 70. 24-144. (canceled)
 145. A method ofdecreasing amyloid plaque load and/or CAA load in the brain of a subjectin need thereof, the method comprising administering a therapeuticallyeffective amount of at least one anti-ApoE antibody, wherein theantibody (a) binds to human ApoE4 with a KD between about 0.1 pM toabout 10 μM, or between about 0.1 pM to about 1 μM, (b) preferentiallybinds recombinant alipidated human ApoE4 as compared to ApoE4 derivedfrom human plasma or human ApoE4 derived from plasma of a transgenicmouse expressing human ApoE4, and (c) binds to human ApoE in amyloidplaque in unfixed brain tissue. 146-147. (canceled)
 148. The method ofclaim 145, wherein the amyloid plaque loads are decreased in thehippocampus and/or in the cortex.
 149. (canceled)
 150. The method ofclaim 145, wherein CAA loads are decreased in the penetrating arterioleson the surface of the cerebral cortex and/or the and leptomeningealarterioles on the surface of the cerebral cortex. 151-165. (canceled)166. A method of treating at least one Aß plaque-associated symptom orat least one CAA-associated symptom in a subject in need thereof, themethod comprising administering a therapeutically effective amount of atleast one anti-ApoE antibody, wherein the antibody (a) binds to humanApoE4 with a KD between about 0.1 pM to about 10 μM, or between about0.1 pM to about 1 μM, (b) preferentially binds recombinant alipidatedhuman ApoE4 as compared to ApoE4 derived from human plasma or humanApoE4 derived from plasma of a transgenic mouse expressing human ApoE4,and (c) binds to human ApoE in amyloid plaque in unfixed brain tissue.167. (canceled)
 168. The method of claim 166, wherein the Aßplaque-associated symptom or the CAA-associated symptom is selected fromneuronal degeneration, impaired cognitive function, altered behavior,abnormal language function, emotional dysregulation, seizures, impairednervous system structure, impaired nervous system function, an increasedrisk of development of Alzheimer's disease, or an increased risk ofdevelopment of cerebral amyloid angiopathy. 169-218. (canceled)